Organic compounds

ABSTRACT

The present invention relates to a compound of the formula I 
                         
wherein R1, R2, R3, R4 and R5 are as defined in the specification, for use in the diagnostic and therapeutic treatment of a warm-blooded animal, especially for the treatment of a disease (=disorder) that depends on activity of renin; the use of a compound of that class for the preparation of a pharmaceutical formulation for the treatment of a disease that depends on activity of renin; the use of a compound of that class in the treatment of a disease that depends on activity of renin; pharmaceutical formulations a compound of that class; a method of treatment comprising administering a compound of that class and a method for its manufacture.

This application is a continuation of U.S. application Ser. No.12/146073 which claims benefit under 35 U.S.C. §119(a)-(d) or (f) or365(b) of EP Applications No. 07111209.8, filed Jun. 28, 2007, and07012412.8, filed Jun. 25, 2007, the contents of which are incorporatedherein by reference in their entirety.

The invention relates to 3,5-substituted piperidine compounds of formulaI, these compounds for use in the diagnostic and therapeutic treatmentof a warm-blooded animal, especially for the treatment of a disease(=disorder) that depends on activity of renin; the use of a compound ofthat class for the preparation of a pharmaceutical formulation for thetreatment of a disease that depends on activity of renin; the use of acompound of that class in the treatment of a disease that depends onactivity of renin; pharmaceutical formulations comprising said3,5-substituted piperidine compound; a method of treatment comprisingadministering said 3,5-substituted piperidine compound and a method forthe manufacture of said 3,5-substituted piperidine compounds.

We have recently described novel 3,5-substituted piperidines which areuseful as renin inhibitors (see PCT/EP06/012581). Although thesecompounds are suitable and effective for this purpose, there is acontinued need to develop renin inhibitors with a further improvedpharmacokinetic profile whilst at the same time achieving a good potencyand safety profile. In particular, the provision of renin inhibitorswith enhanced bioavailability is of therapeutic advantage.Bioavailability is an important factor limiting the therapeuticapplications of bioactive compounds. The object of the present inventionwas thus to provide novel potent renin inhibitors with enhancedbioavailability.

The present invention relates to a compound of the formula I

wherein

-   R1 is C₁₋₇alkyl, which is optionally substituted by one, two or    three substituents selected form the group consisting of hydroxyl,    halo and C₁-C₇-alkoxy;-   R2 is hydrogen, C₁₋₇alkyl, C₁-C₇-alkoxy, halo-C₁-C₇-alkyl,    halo-C₁-C₇-alkoxy or C₁-C₇-alkoxy-C₁-C₇-alkoxy;-   R3 is C₁₋₇alkyl or C₃₋₈cycloalkyl;-   R4 is hydrogen or hydroxyl; and-   R5 is C₁₋₇alkyl or C₃₋₈cycloalkyl;-   or a salt thereof.

In one preferred embodiment the invention relates to a compound of theformula I wherein

-   R1 is C₁₋₇alkyl, which is optionally substituted by one, two or    three substituents selected form the group consisting of hydroxyl,    halo and C₁-C₇-alkoxy;-   R2 is hydrogen, C₁₋₇alkyl, C₁-C₇-alkoxy, halo-C₁-C₇-alkyl or    halo-C₁-C₇-alkoxy;-   R3 is C₁₋₇alkyl or C₃₋₈cycloalkyl;-   R4 is hydrogen or hydroxyl; and-   R5 is C₁₋₇alkyl or C₃₋₈cycloalkyl;-   or a salt thereof.

The present invention also relates to a compound of the formula II

wherein

-   R1 is C₁₋₇alkyl, which is optionally substituted by one, two or    three substituents selected form the group consisting of hydroxyl,    halo and C₁-C₇-alkoxy;-   R2 is hydrogen, C₁₋₇alkyl, C₁-C₇-alkoxy, halo-C₁-C₇-alkyl,    halo-C₁-C₇-alkoxy or C₁-C₇-alkoxy-C₁-C₇-alkoxy;-   R3 is C₁₋₇alkyl or C₃₋₈cycloalkyl;-   R4 is hydrogen or hydroxyl; and-   R5 is C₁₋₇alkyl or C₃₋₈cycloalkyl;-   or a salt thereof.

In another preferred embodiment the invention relates to a compound ofthe formula II wherein

-   R1 is C₁₋₇alkyl, which is optionally substituted by one, two or    three substituents selected form the group consisting of hydroxyl,    halo and C₁-C₇-alkoxy;-   R2 is hydrogen, C₁₋₇alkyl, C₁-C₇-alkoxy, halo-C₁-C₇-alkyl or    halo-C₁-C₇-alkoxy;-   R3 is C₁₋₇alkyl or C₃₋₈cycloalkyl;-   R4 is hydrogen or hydroxyl; and-   R5 is C₁₋₇alkyl or C₃₋₈cycloalkyl;-   or a salt thereof.

The compounds of the formula II are preferred embodiments of the formulaI wherein R4 is hydrogen.

The present invention also relates to a compound of the formula III

R1 is C₁₋₇alkyl, which is optionally substituted by one, two or threesubstituents selected form the group consisting of hydroxyl, halo andC₁-C₇-alkoxy;

R2 is hydrogen, C₁₋₇alkyl, C₁-C₇-alkoxy, halo-C₁-C₇-alkyl,halo-C₁-C₇-alkoxy or C₁-C₇-alkoxy-C₁-C₇-alkoxy;

R3 is C₁₋₇alkyl or C₃₋₈cycloalkyl;

R4 is hydrogen or hydroxyl; and

R5 is C₁₋₇alkyl or C₃₋₈cycloalkyl;

or a salt thereof.

The compounds of the formula III are preferred embodiments of theformula I wherein R4 is hydroxyl.

The compounds of the present invention exhibit inhibitory activity onthe natural enzyme renin. Thus, compounds of formula I may be employedfor the treatment (this term also including prophylaxis) of one or moredisorders or diseases especially selected from the diseases given indetail below, especially as far as these diseases can be modulated (moreespecially beneficially influenced) by renin inhibition. Listed beloware definitions of various terms used to describe the compounds of thepresent invention as well as their use and synthesis, starting materialsand intermediates and the like. These definitions, either by replacingone, more than one or all general expressions or symbols used in thepresent disclosure and thus yielding preferred embodiments of theinvention, preferably apply to the terms as they are used throughout thespecification unless they are otherwise limited in specific instanceseither individually or as part of a larger group.

Alkyl is C₁-C₇-alkyl, more preferably C₁-C₄-alkyl, that isstraight-chained or branched (one or, where appropriate, more times).The term C₁-C₇alkyl defines a moiety with up to and including maximally7, especially up to and including maximally 4, carbon atoms, said moietybeing branched (one or more times) or straight-chained and bound via aterminal or a non-terminal carbon. C₁-C₇-alkyl, for example, isn-pentyl, n-hexyl or n-heptyl or preferably C₁-C₄-alkyl, especially asmethyl, ethyl, n-propyl, sec-propyl, n-butyl, isobutyl, sec-butyl ortert-butyl.

Halo or halogen is preferably fluoro, chloro, bromo or iodo, mostpreferably fluoro, chloro or bromo; where halo is mentioned, this canmean that one or more (e.g. up to three) halogen atoms are present inmoieties such as halo-C₁-C₇-alkyl, halo-C₁-C₇-alkoxy and the like (e.g.trifluoromethyl).

C₃-C₈-cycloalkyl is preferably mono- or bicyclic, more preferablymonocyclic, cycloalkyl, which may include one or more double and/ortriple bonds, C₃-C₆-cycloalkyl is preferred.

C₁-C₇-alkoxy is, for example, C₁-C₄-alkoxy and may be linear orbranched. Examples are methoxy, ethoxy, n- and i-propyloxy, n-, i- andt-butyloxy, pentyloxy and hexyloxy. C₁-C₄ alkoxy is preferred.

Aryl preferably is a mono- or bicyclic aryl with 6 to 22 carbon atoms,especially phenyl, indenyl, indanyl or naphthyl, in particular phenyl,

Alkenyl may be linear or branched alkyl containing a double bond andcomprising preferably 2 to 12 C atoms, 2 to 8 C atoms being especiallypreferred. Particularly preferred is a linear C₂₋₄ alkenyl. Especiallypreferred is allyl.

Bonds with the asterisk (*) denote point of binding to the rest of themolecule.

In all definitions above and below the person having skill in the artwill, without undue experimentation or effort, be able to recognizewhich are especially relevant (e.g. those that if present providecompounds that are sufficiently stable for the manufacture ofpharmaceuticals, e.g. having a half-life of more than 30 seconds,preferably of more than a week) and thus are preferably encompassed bythe present claims and that only chemically feasible bonds andsubstitutions (e.g. in the case of double or triple bonds, hydrogencarrying amino or hydroxy groups and the like can be avoided in order toavoid tautomerism) are encompassed, as well as tautomeric forms wherepresent, especially in equilibrium. For example, preferably, for reasonsof stability or chemical feasibility, directly vicinal atoms in chainspreferably are not selected from oxy plus oxy, thio plus oxy, oxy plusthio or thio plus thio, except where ring systems or the like arepresent that are sufficiently stable. Substitutents binding via an O(e.g. in C₁-C₇-alkoxy) or S that is part of them are preferably notbound to nitrogen e.g. in rings.

Salts are especially the pharmaceutically acceptable salts of compoundsof formula I. They can be formed where salt forming groups, such asbasic or acidic groups, are present that can exist in dissociated format least partially, e.g. in a pH range from 4 to 10 in aqueoussolutions, or can be isolated especially in solid, especiallycrystalline, form.

Such salts are formed, for example, as acid addition salts, preferablywith organic or inorganic acids, from compounds of formula I with abasic nitrogen atom (e.g. imino or amino), especially thepharmaceutically acceptable salts. Suitable inorganic acids are, forexample, halogen acids, such as hydrochloric acid, sulfuric acid, orphosphoric acid. Suitable organic acids are, for example, carboxylic,phosphonic, sulfonic or sulfamic acids, for example acetic acid,propionic acid, lactic acid, fumaric acid, succinic acid, citric acid,amino acids, such as glutamic acid or aspartic acid, maleic acid,hydroxymaleic acid, methylmaleic acid, benzoic acid, methane- orethane-sulfonic acid, ethane-1,2-disulfonic acid, benzenesulfonic acid,2-naphthalenesulfonic acid, 1,5-naphthalene-disulfonic acid,N-cyclohexylsulfamic acid, N-methyl-, N-ethyl- or N-propyl-sulfamicacid, or other organic protonic acids, such as ascorbic acid.

In the presence of negatively charged radicals, such as carboxy orsulfonyl, salts may also be formed with bases, e.g. metal or ammoniumsalts, such as alkali metal or alkaline earth metal salts, for examplesodium, potassium, magnesium or calcium salts, or ammonium salts withammonia or suitable organic amines, such as tertiary monoamines, forexample triethylamine or tri(2-hydroxyethyl)amine, or heterocyclicbases, for example N-ethyl-piperidine or N,N′-dimethylpiperazine.

When a basic group and an acid group are present in the same molecule, acompound of formula I may also form internal salts.

For isolation or purification purposes it is also possible to usepharmaceutically unacceptable salts, for example picrates orperchlorates. For therapeutic use, only pharmaceutically acceptablesalts or free compounds are employed (where applicable comprised inpharmaceutical preparations), and these are therefore preferred.

In view of the close relationship between the compounds in free form andin the form of their salts, including those salts that can be used asintermediates, for example in the purification or identification of thecompounds or salts thereof, any reference to “compounds”, “startingmaterials” and “intermediates” hereinbefore and hereinafter, especiallyto the compound(s) of the formula I or their precursors, is to beunderstood as referring also to one or more salts thereof or a mixtureof a corresponding free compound and one or more salts thereof, each ofwhich is intended to include also any solvate, metabolic precursor suchas ester or amide of the compound of formula I, or salt of any one ormore of these, as appropriate and expedient and if not explicitlymentioned otherwise. Different crystal forms may be obtainable and thenare also included.

Where the plural form is used for compounds, starting materials,intermediates, salts, pharmaceutical preparations, diseases, disordersand the like, this is intended to mean one (preferred) or more singlecompound(s), salt(s), pharmaceutical preparation(s), disease(s),disorder(s) or the like, where the singular or the indefinite article(“a”, “an”) is used, this is intended to include the plural (for examplealso different configuration isomers of the same compound, e.g.enantiomers in racemates or the like) or preferably the singular(“one”).

The compounds of the present invention can possess two or moreasymmetric centers depending on the choice of the substituents. Thepreferred absolute configurations are as indicated herein specifically.However, any possible isolated or pure diastereoisomers, enantiomers orgeometric enantiomers, and mixtures thereof, e.g., mixtures ofenantiomers, such as racemates, are encompassed by the presentinvention.

As described above, the compounds of the present invention areinhibitors of renin activity and, thus, may be employed for thetreatment of hypertension, atherosclerosis, unstable coronary syndrome,congestive heart failure, cardiac hypertrophy, cardiac fibrosis,cardiomyopathy postinfarction, unstable coronary syndrome, diastolicdysfunction, chronic kidney disease, hepatic fibrosis, complicationsresulting from diabetes, such as nephropathy, vasculopathy andneuropathy, diseases of the coronary vessels, restenosis followingangioplasty, raised intra-ocular pressure, glaucoma, abnormal vasculargrowth and/or hyperaldosteronism, and/or further cognitive impairment,Alzheimer's disease, dementia, anxiety states and cognitive disorders,and the like, especially where inhibition of (especially inappropriate)renin activity is required.

“Inappropriate” renin activity preferably relates to a state of awarm-blooded animal, especially a human, where renin shows a reninactivity that is too high in the given situation (e.g. due to one ormore of misregulation, overexpression e.g. due to gene amplification orchromosome rearrangement or infection by microorganisms such as virusthat express an aberrant gene, abnormal activity e.g. leading to anerroneous substrate specificity or a hyperactive renin e.g. produced innormal amounts, too low activity of renin activity product removingpathways, high substrate concentration and/or the like) and/or leads toor supports a renin dependent disease or disorder as mentioned above andbelow, e.g. by too high renin activity. Such inappropriate reninactivity may, for example, comprise a higher than normal activity, orfurther an activity in the normal or even below the normal range which,however, due to preceding, parallel and or subsequent processes, e.g.signaling, regulatory effect on other processes, higher substrate orproduct concentration and the like, leads to direct or indirect supportor maintenance of a disease or disorder, and/or an activity thatsupports the outbreak and/or presence of a disease or disorder in anyother way. The inappropriate activity of renin may or may not bedependent on parallel other mechanisms supporting the disorder ordisease, and/or the prophylactic or therapeutic effect may or mayinclude other mechanisms in addition to inhibition of renin. Therefore“dependent” can be read as “dependent inter alia”, (especially in caseswhere a disease or disorder is really exclusively dependent only onrenin) preferably as “dependent mainly”, more preferably as “dependentessentially only”. A disease dependent on (especially inappropriate)activity of renin may also be one that simply responds to modulation ofrenin activity, especially responding in a beneficial way (e.g. loweringthe blood pressure) in case of renin inhibition.

Where a disease or disorder dependent on (=that “depends on”,“depending”) (especially inappropriate) activity of a renin is mentioned(such in the definition of “use” in the following paragraph and alsoespecially where a compound of the formula I is mentioned for use in thediagnostic or therapeutic treatment which is preferably the treatment ofa disease or disorder dependent on inappropriate renin activity, thisrefers preferably to any one or more diseases or disorders that dependon inappropriate activity of natural renin and/or one or more altered ormutated forms thereof.

Where subsequently or above the term “use” is mentioned (as verb ornoun) (relating to the use of a compound of the formula I or of apharmaceutically acceptable salt thereof, or a method of use thereof),this (if not indicated differently or to be read differently in thecontext) includes any one or more of the following embodiments of theinvention, respectively (if not stated otherwise): the use in thetreatment of a disease or disorder that depends on (especiallyinappropriate) activity of renin, the use for the manufacture ofpharmaceutical compositions for use in the treatment of a disease ordisorder that depends on (especially inappropriate) activity of renin; amethod of use of one or more compounds of the formula I in the treatmentof a disease or disorder that depends on (especially inappropriate)activity of renin; a pharmaceutical preparation comprising one or morecompounds of the formula I for the treatment of a disease or disorderthat depends on (especially inappropriate) activity of renin; and one ormore compounds of the formula I for use in the treatment of a disease ordisorder in a warm-blooded animal, especially a human, preferably adisease that depends on (especially inappropriate) activity of renin; asappropriate and expedient, if not stated otherwise.

The terms “treat”, “treatment” or “therapy” refer to the prophylactic(e.g. delaying or preventing the onset of a disease or disorder) orpreferably therapeutic (including but not limited to preventive, delayof onset and/or progression, palliative, curing, symptom-alleviating,symptom-reducing, patient condition ameliorating, renin-modulatingand/or renin-inhibiting) treatment of said disease(s) or disorder(s),especially of the one or more diseases or disorders mentioned above orbelow.

PREFERRED EMBODIMENTS ACCORDING TO THE INVENTION

The groups of preferred embodiments of the invention mentioned below arenot to be regarded as exclusive, rather, e.g., in order to replacegeneral expressions or symbols with more specific definitions, parts ofthose groups of compounds can be interchanged or exchanged using thedefinitions given above, or omitted, as appropriate, and each of themore specific definitions, independent of any others, may be introducedindependently of or together with one or more other more specificdefinitions for other more general expressions or symbols.

The invention preferably relates to a compound of the formula I, II orIII wherein the moiety R5 is bound in the R configuration oralternatively wherein this moiety is bound in the S configuration.

The invention thus more preferably relates to a compound of the formulaI, II or III as defined herein before or hereinafter which has theconfiguration shown in the following formula IIA or IIB,

or a (preferably pharmaceutically acceptable) salt thereof,or alternatively the configuration shown in the following formula IIIAor IIIB

or a (preferably pharmaceutically acceptable) sat thereof, where informulae IIA, IIB, IIA and IIIB, R1, R2, R3 and R5 are as defined aboveor below for a compound of the formula I.

Alternatively and also more preferably, the invention relates to acompound of the formula I or III as defined herein before or hereinafterwhich has the configuration shown in the following formula III′,

or a (preferably pharmaceutically acceptable) salt thereof, where informulae III′, R1, R2, R3 and R5 are as defined above or below for acompound of the formula I.

Alternatively and also more preferably, the invention relates to acompound of the formula I or III as defined herein before or hereinafterwhich has the configuration shown in the following formula III′A orIII′B,

or a (preferably pharmaceutically acceptable) salt thereof, where informulae III′A and III′B, R1, R2, R3 and R5 are as defined above orbelow for a compound of the formula I.

In a first preferred embodiment, the invention especially relates to acompound of the formulae I, II, III, III′, IIA, IIB, IIIA, IIIB, III′Aor III′B, wherein R1 is a C₁₋₇alkyl, more preferably C₁₋₄-alkyl, whichis optionally substituted by C₁-C₇-alkoxy, more preferably C₁₋₄-alkoxy.In one embodiment, R1 is a C₁₋₇alkyl, which is unsubstituted.Particularly preferred examples for R1 are selected from

In another preferred embodiment, the invention especially relates to acompound of the formulae I, II, III, III′, IIA, IIB, IIIA, IIIB, III′Aor III′B, wherein R2 is hydrogen, C₁-C₇-alkoxy orC₁-C₇-alkoxy-C₁-C₇-alkoxy; more preferably hydrogen, C₁-C₄-alkoxy orC₁-C₄-alkoxy-C₁-C₄-alkoxy. Particularly preferred examples for R2 areselected from hydrogen,

In one preferred embodiment, the invention especially relates to acompound of the formulae I, II, III, III′, IIA, IIB, IIIA, IIIB, III′Aor III′B, wherein R2 is hydrogen or C₁-C₇-alkoxy; more preferablyhydrogen or C₁-C₄-alkoxy. Particularly preferred examples for R2 areselected from hydrogen,

In another preferred embodiment, the invention especially relates to acompound of the formulae I, II, III′, IIA, IIB, IIIA, IIIB, III′A orIII′B, wherein R3 is C₃₋₇alkyl or C₃₋₆-cycloalkyl; more preferablybranched C₄₋₆-alkyl or cyclopropyl.

Particularly preferred examples for R3 are

In one embodiment, R3 is cyclopropyl.

In another preferred embodiment, the invention especially relates to acompound of the formulae I, II, III, III′, IIA, IIB, IIIA, IIIB, III′Aor III′B, wherein R5 is C₁₋₅-alkyl or C₃₋₆-cycloalkyl; more preferablyC₁₋₄alkyl or cyclohexyl. The alkyl can be branched or bonded via anon-terminal C. Particularly preferred examples for R5 are

Particular embodiments of the invention are provided in the Examples—theinvention thus, in a very preferred embodiment, relates to a compound ofthe formula I or a salt thereof, selected from the compounds given inthe Examples, as well as the use thereof according to the invention.

Process of Manufacture

A compound of formula I, or a salt thereof, is prepared especially asdescribed or in analogy to methods described in PCT/EP06/012581, ingeneral by a process comprising:

-   -   a) reacting a compound of the formula IV,

-   -    wherein PG is a protecting group and R4 and R5 are as defined        above, or (preferably) an activated derivative thereof, with a        compound of the formula V,

-   -    wherein R1, R2 and R3 are as defined above; or    -   b) reacting a compound of the formula VI,

-   -    wherein PG is a protecting group and R1, R2, R3 and R4 are as        defined above, or (preferably) an activated derivative thereof,        with a compound of the formula VII,

wherein R5 is as defined above;

-   and, if desired, subsequent to any one or more of the processes    mentioned above converting an obtainable compound of the formula I    or a protected form thereof into a different compound of the formula    I, converting a salt of an obtainable compound of formula I into the    free compound or a different salt, converting an obtainable free    compound of formula I into a salt thereof, and/or separating an    obtainable mixture of isomers of a compound of formula I into    individual isomers;-   where in any of the starting materials, in addition to specific    protecting groups mentioned, further protecting groups may be    present, and any protecting groups or bound resins are removed at an    appropriate stage in order to obtain a corresponding compound of the    formula I, or a salt thereof.

Alternatively, a compound of formula I, or a salt thereof, is preparedin general by a process comprising:

reacting a compound of formula (IXa)

wherein PG is a protecting group, R4 is as defined above and R6 isunsubstituted or substituted alkyl or alkenyl, preferably C₁-C₄ alkyl,or (preferably) an activated derivative thereof, with a compound of theformula V,

wherein R1, R2 and R3 are as defined above; to obtain the amide offormula Xa

which is subjected to hydrolysis of the ester moiety to obtain acompound of formula XIa

which compound or (preferably) an activated derivative thereof, can bein turn reacted with a compound of the formula VII,

wherein R5 is as defined above, to obtain a compound of formula XII

and, if desired, subsequent to any one or more of the processesmentioned above converting an obtainable compound of the formula I or aprotected form thereof into a different compound of the formula I,converting a salt of an obtainable compound of formula I into the freecompound or a different salt, converting an obtainable free compound offormula I into a salt thereof, and/or separating an obtainable mixtureof isomers of a compound of formula I into individual isomers;

-   where in any of the starting materials, in addition to specific    protecting groups mentioned, further protecting groups may be    present, and any protecting groups or bound resins are removed at an    appropriate stage in order to obtain a corresponding compound of the    formula I, or a salt thereof.

In addition, a compound of formula I, or a salt thereof, can be preparedin general by a process comprising:

reacting a compound of formula (IXb)

wherein PG is a protecting group, R4 is as defined above and R6 isunsubstituted or substituted alkyl or alkenyl, preferably C₁-C₄ alkyl,or (preferably) an activated derivative thereof, with a compound of theformula VII,

wherein R5 is as defined above, to obtain the amide of formula Xb

which is subjected to hydrolysis of the ester moiety to obtain acompound of formula XIb

which compound or (preferably) an activated derivative thereof, can bein turn reacted with a compound of the formula V,

wherein R1, R2 and R3 are as defined above, to obtain a compound offormula XII

and, if desired, subsequent to any one or more of the processesmentioned above converting an obtainable compound of the formula I or aprotected form thereof into a different compound of the formula I,converting a salt of an obtainable compound of formula I into the freecompound or a different salt, converting an obtainable free compound offormula I into a salt thereof, and/or separating an obtainable mixtureof isomers of a compound of formula I into individual isomers;

-   where in any of the starting materials, in addition to specific    protecting groups mentioned, further protecting groups may be    present, and any protecting groups or bound resins are removed at an    appropriate stage in order to obtain a corresponding compound of the    formula I, or a salt thereof.

In the case of a compound of the formula IV, VI, IXa, IXb, XIa or XIb,an activated derivative thereof, is the corresponding compound whereinthe OH group of the carboxyl is preferably replaced by a leaving group,such as halo, e.g. chloro, bromo or iodo, or organic sulfonyloxy, suchas tosyloxy or methanesulfonyloxy. The reaction of such activatedderivative thereof with a compound of the formula V or VII thenpreferably takes under standard conditions for nucleophilicsubstitution. In a preferred embodiment, activated derivatives include,for example, acyl halides, anhydrides, and activated esters. Anactivated ester is one which is known to one skilled in the art, forexample N-succinimide. An activated derivative of a carboxylic acid offormula IV, VI, IXa or IXb is, for example, the correspondingintermediate compound formed upon reaction of said acid with EDCI, BopClor TcBocCl. In another preferred embodiment, an activated derivative ofcompounds of formula IV, VI, IXa, IXb, XIa or XIb is the correspondingderivative formed under Vilsmeier reaction conditions.

Moreover, a process for the manufacture of a compound of the formula I,or a pharmaceutically acceptable salt thereof, can comprise reacting acompound of the formula VIII

wherein PG is a protecting group, with an alcohol R6OH, wherein R6 isunsubstituted or substituted alkyl or alkenyl, preferably C₁-C₄ alkyl,in the presence of a chiral amine catalyst, to obtain a compound offormula VIII′a or VIII′b

The nature of the amine governs the position of the ester formation andthus the stereoselectivity. Preferred examples of the chiral aminecatalyst are chiral tertiary amines, more preferably cinchona alkaloids,such as quinidine and quinine, most preferably modified cinchonaalkaloids. Examples of such modified cinchona alkaloids are given below.For a more detailed description, reference is made to Tian, S.-K.; Chen,Y.; Hang, J.; Tang, L.; McDiad, P.; Deng, L. Acc. Chem. Res. 2004, 37,621-631 and references cited therein.

Abbreviations: DHQD-CLB Hydroquinidine 4-chlorobenzoate DHQ-CLBHydroquinine 4-chlorobenzoate DHQD-MEQ Hydroquinidine4-methyl-2-quinolyl ether DHQ-MEQ Hydroquinine 4-methyl-2-quinolyl etherDHQD-PHN Hydroquinidine 9-O-(9′-phenanthryl)ether DHQ-PHN Hydroquinine9-O-(9′-phenanthryl)ether (DHQD)2PYR Hydroquinidine2,5-diphenyl-4,6-pyrimidinediyl diether (DHQ)2PYR Hydroquinine2,5-diphenyl-4,6-pyrimidinediyl diether (DHQD)2PHAL Hydroquinidine1,4-phthalazinediyl diether (DHQ)2PHAL Hydroquinine 1,4-phthalazinediyldiether (DHQD)2AQN Hydroquinidine anthraquinone-1,4-diyl diether(DHQ)2AQN Hydroquinine anthraquinone-1,4-diyl diether

Preferred amines are DHQ, such as (DHQ)₂AQN, and DHQD, such as(DHQD)₂AQN. For reaction conditions, reference is made again to Tian,S.-K. et al. Specifically, the chiral amine catalyst is typicallyemployed in a below-equimolar amount, preferably below 50 mol %, such as5 to 40 mol %, more preferably 10 to 35 mol %, most preferably 30 mol %.

The alcohol R6OH is a suitable alcohol for esterification on a compoundof formula VIII and hydrolysis in the presence of an amide, such asunsubstituted or substituted alkyl or alkenyl, preferably C₁-C₄ alkyl,whereby substituted alkyl is preferably selected from halo, aryl orsubstituted alkyl, in particular fluoro or phenyl. Examples ofsubstituted alkyl include trifluoromethyl, difluoromethyl,difluoroethyl, fluoromethyl, fluoroethyl or benzyl. Most preferably,R6OH is methanoll.

Reaction solvents can be chosen as described below, in particularpreferred are ethereal solvents such as diethyl ether or tetrahydrofuran(THF) or mixtures of these such as diethyl ether:THF (4:1) or (3:1). Thereaction temperature can be chosen so as to bring the reaction tocompletion in an efficient manner while at the same time suppress theformation of unwanted side products as much as possible. Typicalreaction temperatures are −100 to 20° C., such as −80 to 10° C.,preferably −40 to 0° C.

Separation of the products VIII′a or VIII′b can be achieved by suitablerecrystallization techniques known in the art. For example, reference ismade to the recrystallization methods employed in Park, J.-S.; Yeom,C.-E.; Choi, S. H.; Ahn, Y. S.; Ro, S.; Jeom, Y. H.; Shin, D.-K.; Kim,B. M. Tetrahedron Lett. 2003, 44, 1611-1614 and the reference citedtherein. Thus, chiral amines such as α-phenylethylamine can be employed.

Scheme 1a and 1b show two preferred methods for preparing a compound offormula II. It should be noted that the brief description on each of thearrows for each conversion has been added for illustration purposes onlyand should not be regarded as limiting with respect to the sequence oreach individual step.

Ester hydrolysis and deprotection can be effected according to methodswell known in the art, for example as described in reference books suchas Richard C. Larock, “Comprehensive Organic Transformations: A Guide toFunctional Group Preparations”, Second Edition, Wiley-VCH Verlag GmbH,2000 or in T. W. Greene and P. G. M. Wuts, “Protective Groups in OrganicSynthesis”, Third edition, Wiley, New York 1999; see also the referencemade below with respect to protection and deprotection methods.

The present invention includes all pharmaceutically acceptableisotopically-labeled compounds of formula (I) wherein one or more atomsare replaced by atoms having the same atomic number, but an atomic massor mass number different from the atomic mass or mass number usuallyfound in nature.

Examples of isotopes suitable for inclusion in the compounds of theinvention comprises isotopes of hydrogen, such as ²H and ³H, carbon,such as ¹¹C, ¹³C and ¹⁴C, chlorine, such as ³⁶Cl, fluorine, such as ¹⁸F,iodine, such as ¹²³I and ¹²⁵I, nitrogen, such as ¹³N and ¹⁵N, oxygen,such as ¹⁵O, ¹⁷O and ¹⁸O, phosphorus, such as ³²P, and sulphur, such as³⁵S.

Substitution with heavier isotopes such as deuterium, i.e. ²H, mayafford certain therapeutic advantages resulting from greater metabolicstability, for example, increased in vivo half-life or reduced dosagerequirements, and hence may be preferred in some circumstances.

Isotopically-labeled compounds of formula (I) can generally be preparedby conventional techniques known to those skilled in the art or byprocesses analogous to those described in the accompanying Examples andPreparations Sections using an appropriate isotopically-labeled reagentin place of the non-labeled reagent previously employed.

Starting Materials

In the subsequent description of starting materials (this term includingalso intermediates) and their synthesis, R1, R2, R3, R4, R5, R6, R7, R8and PG have the meanings given above or in the Examples for therespective starting materials or intermediates, if not indicatedotherwise directly or by the context. Protecting groups, if notspecifically mentioned, can be introduced and removed at appropriatesteps in order to prevent functional groups, the reaction of which isnot desired in the corresponding reaction step or steps, employingprotecting groups, methods for their introduction and their removal areas described above or below. The person skilled in the art will readilybe able to decide whether and which protecting groups are useful orrequired.

Where for any of the starting materials isomers (e.g. diastereomers,enantiomers) are present, they can be separated according to standardprocedures at appropriate stages. Other starting materials are known inthe art, commercially available, and/or they can be found in or derivedanalogously from the Examples.

General Process Conditions

The following applies in general (where possible) to all processesmentioned hereinbefore and hereinafter, while reaction conditionsspecifically mentioned above or below are preferred:

In any of the reactions mentioned hereinbefore and hereinafter,protecting groups may be used where appropriate or desired, even if thisis not mentioned specifically, to protect functional groups that are notintended to take part in a given reaction, and they can be introducedand/or removed at appropriate or desired stages. Reactions comprisingthe use of protecting groups are therefore included as possible whereverreactions without specific mentioning of protection and/or deprotectionare described in this specification.

Within the scope of this disclosure only a readily removable group thatis not a constituent of the particular desired end product of formula Iis designated a “protecting group” or PG, unless the context indicatesotherwise. The protection of functional groups by such protectinggroups, the protecting groups themselves, and the reactions appropriatefor their introduction and removal are described for example in standardreference works, such as J. F. W. McOmie, “Protective Groups in OrganicChemistry”, Plenum Press, London and New York 1973, in T. W. Greene andP. G. M. Wuts, “Protective Groups in Organic Synthesis”, Third edition,Wiley, New York 1999, in “The Peptides”; Volume 3 (editors: E. Gross andJ. Meienhofer), Academic Press, London and New York 1981, in “Methodender organischen Chemie” (Methods of Organic Chemistry), Houben Weyl, 4thedition, Volume 15/I, Georg Thieme Verlag, Stuttgart 1974, in H.-D.Jakubke and H. Jeschkeit, “Aminosäuren, Peptide, Proteine” (Amino acids,Peptides, Proteins), Verlag Chemie, Weinheim, Deerfield Beach, and Basel1982, and in Jochen Lehmann, “Chemie der Kohlenhydrate: Monosaccharideund Derivate” (Chemistry of Carbohydrates: Monosaccharides andDerivatives), Georg Thieme Verlag, Stuttgart 1974. A characteristic ofprotecting groups is that they can be removed readily (i.e. without theoccurrence of undesired secondary reactions) for example by solvolysis,reduction, photolysis or alternatively under physiological conditions(e.g. by enzymatic cleavage).

Examples of the protecting group or PG, in particular for nitrogen suchas the piperidine nitrogen of the compounds described herein arealkoxycarbonyl, sulfonyl and acyl groups. Preferred protecting groupscomprise, for example, (i) C₁-C₂-alkyl that is mono-, di- ortrisubstituted by phenyl, such as benzyl, (or) benzhydryl or trityl,wherein the phenyl ring is unsubstituted or substituted by one or more,e.g. two or three, residues e.g. those selected from the groupconsisting of C₁-C₇-alkyl, hydroxy, C₁-C₇-alkoxy, halogen, nitro, cyano,and CF₃; phenyl-C₁-C₂-alkoxycarbonyl; and allyl or cinnamyl. Especiallypreferred are lower (e.g. C₁-C₇) alkoxycarbonyl, such astert-butoxycarbonyl or benzyloxycarbonyl; benzyloxycarbonyl (Cbz),9-fluorenylmethyloxycarbonyl (Fmoc), benzyloxymethyl (BOM),pivaloyl-oxy-methyl (POM), trichloroethxoycarbonyl (Troc),1-adamantyloxycarbonxyl (Adoc), but can also be benzyl, cumyl,benzhydryl, trityl, allyl, alloc (allyloxycarbonyl). The protectinggroup can also be silyl, like trialklysilyl, especially trimethylsilyl,tert-butyl-dimethylsilyl, triethylsilyl, triisopropylsilyl,trimethylsilyethoxymethyl (SEM), and can also be substituted sulfonyl orsubstituted sulfenyl. Most preferred is lower (e.g. C₁-C₇)alkoxycarbonyl, such as tert-butoxycarbonyl. The protecting group mayalso be a sulfonyl group, preferably an aryl sulfonyl group such as asubstituted or unsubstituted phenyl sulfonyl group. In this case,phenyl, if substituted, may be mono-, di- or tri-substituted, preferablymono- or di-substituted with a suitable substituent such as C₁-C₇-alkyl,C₁-C₇-alkoxy, halo-C₁-C₇-alkyl, halo-C₁-C₇-alkoxy, halo, hydroxy, nitro,cyano, more preferably nitro or methyl. Particularly preferred examplesof the sulfonyl protecting group are 2,4-dinitrophenylsulfonyl,4-nitrophenyl sulfonyl, 2-nitrophenyl sulfonyl and 4-methylphenylsulfonyl.

All the above-mentioned process steps can be carried out under reactionconditions that are known per se, preferably those mentionedspecifically, in the absence or, customarily, in the presence ofsolvents or diluents, preferably solvents or diluents that are inerttowards the reagents used and dissolve them, in the absence or presenceof catalysts, condensation or neutralizing agents, for example ionexchangers, such as cation exchangers, e.g. in the H⁺ form, depending onthe nature of the reaction and/or of the reactants at reduced, normal orelevated temperature, for example in a temperature range of from about−100° C. to about 190° C., preferably from approximately −80° C. toapproximately 150° C., for example at from −80 to −60° C., at roomtemperature, at from −20 to 40° C. or at reflux temperature, underatmospheric pressure or in a closed vessel, where appropriate underpressure, and/or in an inert atmosphere, for example under an argon ornitrogen atmosphere. The solvents from which those solvents that aresuitable for any particular reaction may be selected include thosementioned specifically or, for example, water, esters, such as loweralkyl-lower alkanoates, for example ethyl acetate, ethers, such asaliphatic ethers, for example diethyl ether, or cyclic ethers, forexample tetrahydrofurane or dioxane, liquid aromatic hydrocarbons, suchas benzene or toluene, alcohols, such as methanol, ethanol or 1- or2-propanol, nitriles, such as acetonitrile, halogenated hydrocarbons,e.g. as methylene chloride or chloroform, acid amides, such asdimethylformamide or dimethyl acetamide, bases, such as heterocyclicnitrogen bases, for example pyridine or N-methylpyrrolidin-2-one,carboxylic acid anhydrides, such as lower alkanoic acid anhydrides, forexample acetic anhydride, cyclic, linear or branched hydrocarbons, suchas cyclohexane, hexane or isopentane, or mixtures of these, for exampleaqueous solutions, unless otherwise indicated in the description of theprocesses. Such solvent mixtures may also be used in working up, forexample by chromatography or partitioning.

The invention relates also to those forms of the processes in which acompound obtainable as intermediate at any stage of the process is usedas starting material and the remaining process steps are carried out, orin which a starting material is formed under the reaction conditions oris used in the form of a derivative, for example in protected form or inthe form of a salt, or a compound obtainable by the process according tothe invention is produced under the process conditions and processedfurther in situ. In the processes of the present invention thosestarting materials are preferably used which result in compounds offormula I described as being preferred. Special preference is given toreaction conditions that are identical or analogous to those mentionedin the Examples. The invention relates also to novel starting compoundsand intermediates described herein, especially those leading to novelcompounds of the formula I or compounds of the formula I mentioned aspreferred herein.

Pharmaceutical Use, Pharmaceutical Preparations and Methods

As described above, the compounds of the formula I are inhibitors ofrenin activity and, thus, may be of use for the treatment ofhypertension, atherosclerosis, unstable coronary syndrome, congestiveheart failure, cardiac hypertrophy, cardiac fibrosis, cardiomyopathypostinfarction, unstable coronary syndrome, diastolic dysfunction,chronic kidney disease, hepatic fibrosis, complications resulting fromdiabetes, such as nephropathy, vasculopathy and neuropathy, diseases ofthe coronary vessels, restenosis following angioplasty, raisedintra-ocular pressure, glaucoma, abnormal vascular growth and/orhyperaldosteronism, and/or further cognitive impairment, alzheimers,dementia, anxiety states and cognitive disorders, and the like.Hypertension, at least as one component of the disease to be treated, isespecially preferred, meaning that hypertension alone or in combinationwith one or more (especially of the mentioned) other diseases may betreated (prophylactically and/or therapeutically).

The present invention further provides pharmaceutical compositionscomprising a therapeutically effective amount of a pharmacologicallyactive compound of the formula I, alone or in combination with one ormore pharmaceutically acceptable carriers.

The pharmaceutical compositions according to the present invention arethose suitable for enteral, such as oral or rectal, transdermal andparenteral administration to mammals, including man, to inhibit reninactivity, and for the treatment of conditions associated with(especially inappropriate) renin activity. Such conditions includehypertension, atherosclerosis, unstable coronary syndrome, congestiveheart failure, cardiac hypertrophy, cardiac fibrosis, cardiomyopathypostinfarction, unstable coronary syndrome, diastolic dysfunction,chronic kidney disease, hepatic fibrosis, complications resulting fromdiabetes, such as nephropathy, vasculopathy and neuropathy, diseases ofthe coronary vessels, restenosis following angioplasty, raisedintra-ocular pressure, glaucoma, abnormal vascular growth and/orhyperaldosteronism, and/or further cognitive impairment, alzheimers,dementia, anxiety states and cognitive disorders and the like.Especially preferred is a disease which comprises hypertension, moreespecially hypertension itself, where treatment with a pharmaceuticalcomposition or the use of a compound of the formula I for its synthesisis useful prophylactically and/or (preferably) therapeutically.

Thus, the pharmacologically active compounds of the formula I may beemployed in the manufacture of pharmaceutical compositions comprising aneffective amount thereof in conjunction or admixture with excipients orcarriers suitable for either enteral or parenteral application.Preferred are tablets and gelatin capsules comprising the activeingredient together with:

-   a) diluents, e.g., lactose, dextrose, sucrose, mannitol, sorbitol,    cellulose and/or glycine;-   b) lubricants, e.g., silica, talcum, stearic acid, its magnesium or    calcium salt and/or polyethyleneglycol; for tablets also-   c) binders, e.g., magnesium aluminum silicate, starch paste,    gelatin, tragacanth, methylcellulose, sodium carboxymethylcellulose    and or polyvinylpyrrolidone; if desired-   d) disintegrants, e.g., starches, agar, alginic acid or its sodium    salt, or effervescent mixtures; and/or-   e) absorbants, colorants, flavors and sweeteners.

Injectable compositions are preferably aqueous isotonic solutions orsuspensions, and suppositories are advantageously prepared from fattyemulsions or suspensions.

Said compositions may be sterilized and/or contain adjuvants, such aspreserving, stabilizing, wetting or emulsifying agents, solutionpromoters, salts for regulating the osmotic pressure and/or buffers. Inaddition, they may also contain other therapeutically valuablesubstances. Said compositions are prepared according to conventionalmixing, granulating or coating methods, respectively, and contain about0.1-75%, preferably about 1-50%, of the active ingredient.

Suitable formulations for transdermal application include atherapeutically effective amount of a compound of the invention withcarrier. Advantageous carriers include absorbable pharmacologicallyacceptable solvents to assist passage through the skin of the host.Characteristically, transdermal devices are in the form of a bandagecomprising a backing member, a reservoir containing the compoundoptionally with carriers, optionally a rate controlling barrier todeliver the compound of the skin of the host at a controlled andpre-determined rate over a prolonged period of time, and means to securethe device to the skin.

Accordingly, the present invention provides pharmaceutical compositionsas described above for the treatment of conditions mediated by reninactivity, preferably, hypertension, atherosclerosis, unstable coronarysyndrome, congestive heart failure, cardiac hypertrophy, cardiacfibrosis, cardiomyopathy postinfarction, unstable coronary syndrome,diastolic dysfunction, chronic kidney disease, hepatic fibrosis,complications resulting from diabetes, such as nephropathy, vasculopathyand neuropathy, diseases of the coronary vessels, restenosis followingangioplasty, raised intra-ocular pressure, glaucoma, abnormal vasculargrowth and/or hyperaldosteronism, and/or further cognitive impairment,alzheimers, dementia, anxiety states and cognitive disorders, as well asmethods of their use.

The pharmaceutical compositions may contain a therapeutically effectiveamount of a compound of the formula I as defined herein, either alone orin a combination with another therapeutic agent, e.g., each at aneffective therapeutic dose as reported in the art. Such therapeuticagents include:

-   a) antidiabetic agents such as insulin, insulin derivatives and    mimetics; insulin secretagogues such as the sulfonylureas, e.g.,    Glipizide, glyburide and Amaryl; insulinotropic sulfonylurea    receptor ligands such as meglitinides, e.g., nateglinide and    repaglinide; peroxisome proliferator-activated receptor (PPAR)    ligands; protein tyrosine phosphatase-1B (PTP-1B) inhibitors such as    PTP-112; GSK3 (glycogen synthase kinase-3) inhibitors such as    SB-517955, SB-4195052, SB-216763, N,N-57-05441 and N,N-57-05445; RXR    ligands such as GW-0791 and AGN-194204; sodium-dependent glucose    cotransporter inhibitors such as T-1095; glycogen phosphorylase A    inhibitors such as BAY R3401; biguanides such as metformin;    alpha-glucosidase inhibitors such as acarbose; GLP-1 (glucagon like    peptide-1), GLP-1 analogs such as Exendin-4 and GLP-1 mimetics; and    DPPIV (dipeptidyl peptidase IV) inhibitors such as LAF237;-   b) hypolipidemic agents such as 3-hydroxy-3-methyl-glutaryl coenzyme    A (HMG-CoA) reductase inhibitors, e.g., lovastatin, pitavastatin,    simvastatin, pravastatin, cerivastatin, mevastatin, velostatin,    fluvastatin, dalvastatin, atorvastatin, rosuvastatin and rivastatin;    squalene synthase inhibitors; FXR (farnesoid X receptor) and LXR    (liver X receptor) ligands; cholestyramine; fibrates; nicotinic acid    and aspirin;-   c) anti-obesity agents such as orlistat; and-   d) anti-hypertensive agents, e.g., loop diuretics such as ethacrynic    acid, furosemide and torsemide; angiotensin converting enzyme (ACE)    inhibitors such as benazepril, captopril, enalapril, fosinopril,    lisinopril, moexipril, perinodopril, quinapril, ramipril and    trandolapril; inhibitors of the Na-K-ATPase membrane pump such as    digoxin; neutralendopeptidase (NEP) inhibitors; ACE/NEP inhibitors    such as omapatrilat, sampatrilat and fasidotril; angiotensin II    antagonists such as candesartan, eprosartan, irbesartan, losartan,    telmisartan and valsartan, in particular valsartan; β-adrenergic    receptor blockers such as acebutolol, atenolol, betaxolol,    bisoprolol, metoprolol, nadolol, propranolol, sotalol and timolol;    inotropic agents such as digoxin, dobutamine and milrinone; calcium    channel blockers such as amlodipine, bepridil, diltiazem,    felodipine, nicardipine, nimodipine, nifedipine, nisoldipine and    verapamil; aldosterone receptor antagonists; and aldosterone    synthase inhibitors.

Other specific anti-diabetic compounds are described by Patel Mona inExpert Opin Investig Drugs, 2003, 12(4), 623-633, in the FIGS. 1 to 7. Acompound of the formula I may be administered either simultaneously,before or after the other active ingredient, either separately by thesame or different route of administration or together in the samepharmaceutical formulation.

The structure of the therapeutic agents identified by code numbers,generic or trade names may be taken from the actual edition of thestandard compendium “The Merck Index” or from databases, e.g., PatentsInternational (e.g. IMS World Publications).

Accordingly, the present invention provides pharmaceutical products orcompositions comprising a therapeutically effective amount of a compoundof the formula I alone or in combination with a therapeuticallyeffective amount of another therapeutic agent, preferably selected fromanti-diabetics, hypolipidemic agents, anti-obesity agents andanti-hypertensive agents, most preferably from antidiabetics,anti-hypertensive agents and hypolipidemic agents as described above.

The present invention further relates to pharmaceutical compositions asdescribed above for use as a medicament.

The present invention further relates to use of pharmaceuticalcompositions or combinations as described above for the preparation of amedicament for the treatment of conditions mediated by (especiallyinappropriate) renin activity, preferably, hypertension,atherosclerosis, unstable coronary syndrome, congestive heart failure,cardiac hypertrophy, cardiac fibrosis, cardiomyopathy postinfarction,unstable coronary syndrome, diastolic dysfunction, chronic kidneydisease, hepatic fibrosis, complications resulting from diabetes, suchas nephropathy, vasculopathy and neuropathy, diseases of the coronaryvessels, restenosis following angioplasty, raised intra-ocular pressure,glaucoma, abnormal vascular growth and/or hyperaldosteronism, and/orfurther cognitive impairment, alzheimers, dementia, anxiety states andcognitive disorders, and the like.

Thus, the present invention also relates to a compound of formula I foruse as a medicament, to the use of a compound of formula I for thepreparation of a pharmaceutical composition for the prevention and/ortreatment of conditions mediated by (especially inappropriate) reninactivity, and to a pharmaceutical composition for use in conditionsmediated by (especially inappropriate) renin activity comprising acompound of formula I, or a pharmaceutically acceptable salt thereof, inassociation with a pharmaceutically acceptable diluent or carriermaterial.

The present invention further provides a method for the preventionand/or treatment of conditions mediated by (especially inappropriate)renin activity, which comprises administering a therapeuticallyeffective amount of a compound of the formula I to a warm-bloodedanimal, especially a human, in need of such treatment.

A unit dosage for a mammal of about 50-70 kg may contain between about 1mg and 1000 mg, advantageously between about 5-600 mg of the activeingredient. The therapeutically effective dosage of active compound isdependent on the species of warm-blooded animal (especially mammal, moreespecially human), the body weight, age and individual condition, on theform of administration, and on the compound involved.

In accordance with the foregoing the present invention also provides apharmaceutical product comprising a therapeutic combination, e.g., akit, kit of parts, e.g., for use in any method as defined herein,comprising a compound of formula I, or a pharmaceutically acceptablesalt thereof, to be used concomitantly or in sequence with at least onepharmaceutical composition comprising at least another therapeuticagent, preferably selected from anti-diabetic agents, hypolipidemicagents, anti-obesity agents or anti-hypertensive agents. The kit maycomprise instructions for its administration.

Similarly, the present invention provides a kit of parts comprising: (i)a pharmaceutical composition comprising a compound of the formula Iaccording to the invention; and (ii) a pharmaceutical compositioncomprising a compound selected from an anti-diabetic, a hypolipidemicagent, an anti-obesity agent, an anti-hypertensive agent, or apharmaceutically acceptable salt thereof, in the form of two separateunits of the components (i) to (ii).

Likewise, the present invention provides a method as defined abovecomprising co-administration, e.g., concomitantly or in sequence, of atherapeutically effective amount of a compound of formula I, or apharmaceutically acceptable salt thereof, and at least a second drugsubstance, said second drug substance preferably being an anti-diabetic,a hypolipidemic agent, an anti-obesity agent or an anti-hypertensiveagent, e.g., as indicated above.

Preferably, a compound of the invention is administered to a mammal inneed thereof.

Preferably, a compound of the invention is used for the treatment of adisease which responds to a modulation of (especially inappropriate)renin activity, especially one or more of the specific diseasesmentioned above.

Finally, the present invention provides a method or use which comprisesadministering a compound of formula I in combination with atherapeutically effective amount of an anti-diabetic agent, ahypolipidemic agent, an anti-obesity agent or an anti-hypertensiveagent.

Ultimately, the present invention provides a method or use whichcomprises administering a compound of formula I in the form of apharmaceutical composition as described herein.

The above-cited properties are demonstrable in vitro and in vivo testsusing advantageously mammals, e.g., mice, rats, rabbits, dogs, monkeysor isolated organs, tissues and preparations thereof. Said compounds canbe applied in vitro in the form of solutions, e.g., preferably aqueoussolutions, and in vivo either enterally, parenterally, advantageouslyintravenously, e.g., as a suspension or in aqueous solution. Theconcentration level in vitro may range between about 10⁻³ molar and10⁻¹⁰ molar concentrations. A therapeutically effective amount in vivomay range depending on the route of administration, between about 0.001and 500 mg/kg, preferably between about 0.1 and 100 mg/kg.

As described above, the compounds of the present invention haveenzyme-inhibiting properties. In particular, they inhibit the action ofthe natural enzyme renin. Renin passes from the kidneys into the bloodwhere it effects the cleavage of angiotensinogen, releasing thedecapeptide angiotensin I which is then cleaved in the lungs, thekidneys and other organs to form the octapeptide angiotensin II. Theoctapeptide increases blood pressure both directly by arterialvasoconstriction and indirectly by liberating from the adrenal glandsthe sodium-ion-retaining hormone aldosterone, accompanied by an increasein extracellular fluid volume which increase can be attributed to theaction of angiotensin II. Inhibitors of the enzymatic activity of reninlead to a reduction in the formation of angiotensin I, and consequentlya smaller amount of angiotensin II is produced. The reducedconcentration of that active peptide hormone is a direct cause of thehypotensive effect of renin inhibitors.

The action of renin inhibitors may be demonstrated inter aliaexperimentally by means of in vitro tests, the reduction in theformation of angiotensin I being measured in various systems (humanplasma, purified human renin together with synthetic or natural reninsubstrate).

Inter alia the following in vitro tests may be used:

-   1) Recombinant human renin (expressed in Chinese Hamster Ovary cells    and purified using standard methods) at 7.5 nM concentration is    incubated with test compound at various concentrations for 1 h at RT    in 0.1 M Tris-HCl buffer, pH 7.4, containing 0.05 M NaCl, 0.5 mM    EDTA and 0.05% CHAPS. Synthetic peptide substrate    Arg-Glu(EDANS)-Ile-His-Pro-Phe-His-Leu-Val-Ile_His_Thr-Lys(DABCYL)-Arg9    is added to a final concentration of 2 μM and increase in    fluorescence is recorded at an excitation wave-length of 350 nm and    at an emission wave-length of 500 nm in a microplate    spectro-fluorimeter. IC₅₀ values are calculated from percentage of    inhibition of renin activity as a function of test compound    concentration (Fluorescence Resonance Energy Transfer, FRET, assay).    Compounds of the formula I, in this assay, preferably can show IC₅₀    values in the range from 1 nM to 20 μM.-   2) Alternatively, recombinant human renin (expressed in Chinese    Hamster Ovary cells and purified using standard methods) at 0.5 nM    concentration is incubated with test compound at various    concentrations for 2 h at 37° C. in 0.1 M Tris-HCl buffer, pH 7.4,    containing 0.05 M NaCl, 0.5 mM EDTA and 0.05% CHAPS. Synthetic    peptide substrate    Arg-Glu(EDANS)-Ile-His-Pro-Phe-His-Leu-Val-Ile_His_Thr-Lys(DABCYL)-Arg9    is added to a final concentration of 4 μM and increase in    fluorescence is recorded at an excitation wave-length of 340 nm and    at an emission wave-length of 485 nm in a microplate    spectro-fluorimeter. IC50 values are calculated from percentage of    inhibition of renin activity as a function of test compound    concentration (Fluorescence Resonance Energy Transfer, FRET, assay).    Compounds of the formula I, in this assay, preferably can show IC₅₀    values in the range from 1 nM to 20 μM.-   3) In another assay, human plasma spiked with recombinant human    renin (expressed in Chinese Hamster Ovary cells and purified using    standard methods) at 0.8 nM concentration is incubated with test    compound at various concentrations for 2 h at 37° C. in 0.1 M    Tris/HCl pH 7.4 containing 0.05 M NaCl, 0.5 mM EDTA and 0.025% (w/v)    CHAPS. Synthetic peptide substrate    Ac-Ile-His-Pro-Phe-His-Leu-Val-Ile-His-Asn-Lys-[DY-505-X5] is added    to a final concentration of 2.5 μM. The enzyme reaction is stopped    by adding an excess of a blocking inhibitor. The product of the    reaction is separated by capillary electrophoresis and quantified by    spectrophotometric measurement at 505 nM wave-length. IC50 values    are calculated from percentage of inhibition of renin activity as a    function of test compound concentration. Compounds of the formula I,    in this assay, preferably can show IC₅₀ values in the range from 1    nM to 20 μM.-   4) In another assay, recombinant human renin (expressed in Chinese    Hamster Ovary cells and purified using standard methods) at 0.8 nM    concentration is incubated with test compound at various    concentrations for 2 h at 37° C. in 0.1 M Tris/HCl pH 7.4 containing    0.05 M NaCl, 0.5 mM EDTA and 0.025% (w/v) CHAPS. Synthetic peptide    substrate Ac-Ile-His-Pro-Phe-His-Leu-Val-Ile-His-Asn-Lys-[DY-505-X5]    is added to a final concentration of 2.5 μM. The enzyme reaction is    stopped by adding an excess of a blocking inhibitor. The product of    the reaction is separated by capillary electrophoresis and    quantified by spectrophotometric measurement at 505 nM wave-length.    IC50 values are calculated from percentage of inhibition of renin    activity as a function of test compound concentration. Compounds of    the formula I, in this assay, preferably show IC₅₀ values in the    range from 1 nM to 20 μM.

In animals deficient in salt, renin inhibitors bring about a reductionin blood pressure. Human renin may differ from the renin of otherspecies. In order to test inhibitors of human renin, primates, e.g.,marmosets (Callithrix jacchus) may be used, because human renin andprimate renin are substantially homologous in the enzymatically activeregion. Inter alia the following in vivo tests may be used:

Compounds of the formula I can be tested in vivo in primates asdescribed in the literature (see for example by Schnell C R et al.Measurement of blood pressure and heart rate by telemetry in conscious,unrestrained marmosets. Am J Physiol 264 (Heart Circ Physiol 33). 1993:1509-1516; or Schnell C R et al. Measurement of blood pressure, heartrate, body temperature, ECG and activity by telemetry in conscious,unrestrained marmosets. Proceedings of the fifth FELASA symposium:Welfare and Science. Eds BRIGHTON. 1993.

It has been found that the new compounds, beside being potent renininhibitors, also show an improved bioavailability. The bioavailabilityis preferably equal to or higher than 20%, more preferably equal to orhigher than 30%. The bioavailability can be determined as follows.

Pharmacokinetic profiles are investigated in male Sprague-Dawley ratsimplanted with jugular vein catheters. Compounds are administered orallyin 0.5% aqueous methylcellulose solution or intravenously inN-methylpyrrolidinone-PEG200 (10:90, v/v). Typical doses are 6 mg/kgp.o. and 2 mg/kg i.v., respectively. Blood samples are serially takenthrough venous catheters into heparinized tubes at various time pointsuntil 32 h post dose and plasma is separated by centrifugation. Plasmaconcentrations of the compounds described in this invention are measuredby liquid chromatography-tandem mass spectrometry after extraction withacetonitrile.

Pharmacokinetic parameters are calculated by using a non-compartmentalmethod.

Abbreviations Boc tert-butoxycarbonyl BopClbis(2-oxo-3-oxazolidinyl)phosphinic chloride CAN ammonium cerium (IV)nitrate DMF N,N-dimethylformamide DMSO dimethylsulfoxide EDCl.HCl1[3-(dimethylamino)propyl]-3-ethylcarbodiinnide hydrochloride Et ethylEtOAc ethyl acetate h hour(s) HOAT 1-hydroxy-7-azabenzotriazole mLmilliliter Me methyl MS mass spectrometry NMP N-methylpyrrolidinone Phphenyl i-Pr isopropyl RT room temperature TcBocCl2,2,2-trichloro-1,1-dinnethylethyl chloroformate TFA trifluoroaceticacid THF tetrahydrofuran TLC thin layer chromatography t_(R) retentiontime

Temperatures are measured in degrees Celsius. Unless otherwiseindicated, the reactions take place at RT. Unless otherwise indicated,the hydrogenation reactions in the presence of H₂ take place atatmospheric pressure. The microwave irradiation is performed by using a“Biotage Initiator 60” machine.

HPLC Condition-A:

-   Column: CombiScreen ODS-AM, 50×4.6 mm.-   Flow rate: 2.0 mL/min-   Mobile phase: A) TFA/water (0.1/100, v/v), B) TFA/acetonitrile    (0.1/100, v/v)-   Gradient: linear gradient from 5% B to 100% B in 5 min then 100% B    in 2 min-   Detection: UV at 215 nm    HPLC Condition-B:-   Column: ACQUITY UPLC™ BEH C₁₈ 1.7 μm, 50×2.1 mm.-   Flow rate: 0.5 mL/min-   Mobile phase: A) TFA/water (0.1/100, v/v), B) TFA/acetonitrile    (0.1/100, v/v)-   Gradient: 5% B in 0.5 min then linear gradient from 5% B to 100% B    in 1.5 min then 100% B in 1.0 min-   Detection: UV at 215 nm    HPLC Condition-C:-   Column: ACQUITY UPLC™ BEH C₁₈ 1.7 μm, 50×2.1 mm.-   Flow rate: 0.5 mL/min-   Mobile phase: A) TFA/water (0.1/100, v/v), B) TFA/acetonitrile    (0.1/100, v/v)-   Gradient: 5% B in 0.5 min then linear gradient from 5% B to 100% B    in 5.0 min then 100% B in 1.5 min-   Detection: UV at 215 nm-   TLC conditions: R_(f) values for TLC are measured on 5×10 cm TLC    plates, silica gel F₂₅₄, Merck, Darmstadt, Germany.

Methods for preparing compounds of formula I are described in detailbelow. It should be noted that the brief description on each of thearrows for each conversion has been added for illustration purposes onlyand should not be regarded as limiting with respect to the sequence oreach individual step.

A: Pyridine-3,5-dicarboxylic acid dimethyl ester

3,5-Pyridinedicarboxylic acid (1.5 g, 63 mmol) and conc. H₂SO₄ (0.9 mL)in MeOH (15 mL) are heated in a microwave oven at 120° C. for 2 h. Thesolvent is evaporated to give a residue which is partitioned betweenethyl acetate and sat. aq. NaHCO₃. The organic phase is washed withbrine, dried over Na2SO4, filtered and evaporated to give a light yellowsolid.

MS (LC-MS): 196 [M+H]⁺ TLC, Rf (ethyl acetate/hexane 1:1)=0.56.

B: Piperidine-3,5-dicarboxylic acid dimethyl ester

Pyridine-3,5-dicarboxylic acid dimethyl ester (5.3 g, 27 mmol) andRh/PtO₂ (0.5 g) in MeOH (200 mL) are stirred under hydrogen overnight.The resulting mixture is filtered and the solvents are evaporated togive a brown oil. MS (LC-MS): 202 [M+H]⁺

C: Piperidine-1,3,5-tricarboxylic acid 1-tert-butyl ester 3,5-dimethylester

A solution of piperidine-3,5-dicarboxylic acid dimethyl ester (5.4 g,26.8 mmol) in CH₂Cl₂ (55 mL) is treated with Boc₂O (6.4 g, 29.5 mmol)and the reaction is stirred at rt overnight. The reaction is quenchedwith 0.1N aq. HCl and the organic phase is washed with 0.1N aq HCl. Thecombined aqueous phases are extracted 2 times with CH₂Cl₂/MeOH (9/1)before the combined organic phases are dried over Na₂SO₄, filtered andevaporated. The resulting residue is purified by flash chromatography onsilica gel (eluent: CH₂Cl₂/MeOH 95:5) to give the title compound as ayellow solid.

MS (LC-MS): 302 [M+H]⁺.TLC, Rf (CH₂Cl₂/MeOH 95:5)=0.5.

D: Piperidine-1,3,5-tricarboxylic acid 1-tert-butyl ester

To a solution of piperidine-1,3,5-tricarboxylic acid 1-tert-butyl ester3,5-dimethyl ester (6.8 g, 22.5 mmol) in MeOH/water (4:1, 120 mL) isadded K₂CO₃ (9.4 g, 68 mmol).

The reaction mixture is stirred at reflux overnight. The MeOH isevaporated and the residue is extracted with dichloromethane and 1N aq.HCl. The organic phase is dried over Na₂SO₄, filtered and evaporated togive a light yellow solid. MS (LC-MS): 274 [M+H]⁺.

E: 2,4-Dioxo-3-oxa-7-aza-bicyclo[3.3.1]nonane-7-carboxylic acidtert-butyl ester

A suspension of piperidine-1,3,5-tricarboxylic acid 1-tert-butyl ester(1 g, 3.6 mmol) in acetic anhydride (20 mL) is heated at reflux for 2 h.The reaction mixture is evaporated three times with toluene before it isdried under high vacuum at rt overnight to give a yellow solid. MS(LC-MS): 278 [M+Na]⁺.

(3S,5R)-Starting Material-F

To a solution of (3S,5R)-starting material-F (67% ee) (47 g, 162 mmol)in hot EtOH (162 mL) is added (S)-(−)-1-phenylethylamine (20.6 mL, 162mmol) at 80° C. The solution is cooled to r.t. and allowed to stand forovernight, which results in the precipitation of a salt. The salt iscollected by filtration. After repeating the same recrystallizationprocedure in EtOH three times, the resulting salt is dissolved in water,acidified with 5N and 1N HCl, and extracted with AcOEt. The combinedorganic phases are washed with brine, dried over MgSO₄. Concentrationunder reduced pressure gives (3S,5R)-starting material-F: colorlesscrystal; ES-MS: M+H=288: _(B)t_(Ret)=2.67 min. chiral HPLC (column:CHIRALPAH AD-H (0.46 cm×25 cm), eluent: hexane/1-PrOH/0.1% TFA=95/5,flow rate: 0.5 mL/min, detection: UV 210 nm, temperature: rt) t_(R)=37min

(3S,5R)-Starting Material-F (98% ee)

To the solution of starting material-E (401.5 mg, 1.57 mmol) andcommercially available (DHQD)₂AQN (423.6 mg, 0.47 mmol, 95% purity)^(a)in Et₂O (60 mL) and THF (20 mL) under N₂ is added MeOH (0.64 mL, 15.67mmol) at −40° C. After stirring at that temperature for 24 h, sat.citric acid aq. is added. The reaction mixture is extracted with EA. Theorganic phase is washed with brine, dried over Na₂SO₄ and subjected tosilica chromatography to give (3S,5R)-starting material-F in 98% ee as awhite amorphous material ES-MS: M+H-tBu=232; HPLC: _(C)t_(Ret)=2.73 min.chiral HPLC (column: CHIRALPAH AD-H (0.46 cm×25 cm), eluent:hexane/1-PrOH=95/5, flow rate: 0.5 mL/min, detection: UV 210 nm,temperature: rt) t_(R)=33.25 min

(3R,5S)-starting material-F, 35.56 min for (3S,5R)-starting material-F,^(a) Chen, Y.; Tian, S-K.; Deng, Li. J. Am. Chem. Soc. 2000, 122,9542-9543.

(3R,5S)-Starting Material-F

To a solution of (3R,5S)-starting material-F (72% ee) (4.2 g, 14.6 mmol)in hot EtOH (20 mL) is added (R)-1-phenylethylamine (1.79 g, 14.76 mmol)at 70° C. The solution is cooled to rt and allowed to stand for 1 h,which results in the precipitation of a salt. The salt is collected byfiltration. After repeating the same recrystallization procedure threetimes, the resulting salt was dissolved in water, acidified with 1MHClaq and extracted five times with ether. The combined organic phasesare washed with brine, dried with MgSO₄. Concentration under reducedpressure gives (3R,5S)-starting material-F: colorless crystal; ES-MS:M+H=288: _(B)t_(Ret)=2.67 min. chiral HPLC: AD-H, 5% i-PrOH/Hexane, flow0.5 mL/min, 210 nm, t_(Ret)=33 (major), 36 (minor).

(3R,5S)-Starting Material-F (72% ee)

To a solution of starting material-E (200 mg, 0.78 mmol) in THF (10 mL)and ether (30 mL) is added (DHQ)₂AQN (67 mg, 0.08 mmol) and MeOH (0.32mL) at 0° C. under N₂. The resulting mixture is stirred for 5 h at 0° C.After adding 1M HCl aq, the mixture is extracted with EtOAc. The organicphases are washed with brine and dried with MgSO₄. Concentration underreduced pressure and silica gel flash chromatography give(3R,5S)-starting material-F: ES-MS: M+H=288: _(C)t_(Ret)=2.67 min.chiral HPLC: 72% ee, AD-H, 5% i-PrOH/Hexane, flow 0.5 mL/min, 210 nm,t_(Ret)=33 (major), 36 (minor).

(3R,4S,5S)-starting material-G((3R,4S,5S)—N-tert-Butyloxycarbonyl-4-hydroxy-piperidine-3,5-dicarboxylicacid 3-methyl ester)

A mixture of (3R,4S,5S), and (3S,4R,5R),—N-tert-Butyloxycarbonyl-4-hydroxy-piperidine-3,5-dicarboxylic acid3-methyl ester (96% ee) (78 g, 0.26 mol) is dissolved in MeOH (500 mL)and (S)-(−)1-phenylethylamine (33 mL, 0.26 mol) at room temperature isadded. The mixture is stirred for 30 min at room temperature andconcentrated under reduced pressure to provide a salt as a colorlessamorphous. The resulting residue is dissolved in CH₃CN (1.3 L) at 70° C.and stirred vigorously at room temperature for 20 h. A white crystallinesalt is collected by filtration, washed with Et₂O. After repeating thesame recrystallization procedure (dissolution in CH₃CN and stirring atroom temperature) two or three times the resulting salt is dissolved inwater, acidified to pH 3 with 5N and 1N HCl, and extracted with AcOEt.The combined organic phases are dried over Na₂SO₄. Concentration underreduced pressure gives enantiomeric pure (3R,4S,5S)-starting material-Gas a white solid: ES-MS: M+H=304: _(C)t_(Ret)=2.37 min. chiralHPLC: >99.9% ee, AD-H, 7.5% i-PrOH/Hexane/10.1% TFA, flow 0.75 mL/min,210 nm, t_(Ret)=24 min

(3R,4S,5S)-Starting Material-G (96% ee)

To a solution of(3,4-cis-4,5-cis)-N-tert-Butyloxycarbonyl-4-hydroxy-piperidine-3,5-dicarboxylicacid dimethyl ester (18 g, Liang, X.; Lohse, A.; Bols, M. J. Org. Chem.2000, 65, 7432.) in phosphate buffer (0.2M, pH7.5, 540 mL) is addedLipase M (Mucor javanicus) (5.4 g) at room temperature. The reactionmixture is stirred for 6 days at 35° C. and then acidified to pH 3 with5N and 1N HCl. The mixture is extracted with EtOAc, the organic phasesare dried with Na₂SO₄ Concentration under reduced pressure gives(3R,4S,5S)-starting material-G as a white solid: ES-MS: M+H=304:_(C)t_(Ret)=2.37 min. chiral HPLC: 95.8% ee, AD-H, 7.5%i-PrOH/Hexane/0.1% TFA, flow 0.75 mL/min, 210 nm, t_(Ret)=24 min(3R,4S,5S), 26 min (3S,4R,5R).

EXAMPLE 1

A mixture of Intermediate 1.1 (70 mg, 0.125 mmol) in 4N HCl in dioxane(2 mL) is stirred at room temperature. After stirring for 1 h, thereaction mixture is concentrated in vacuo to give Example 1: ES-MS:M+H=459: _(C)t_(Ret)=3.26 min.

Intermediate 1.1

To a solution of Intermediate 1.4 (27 mg, 0.150 mmol) and Intermediate1.9 (50 mg, 0.125 mmol) in CH₂Cl₂ are added BopCl (47 mg, 0.188) andEt₃N (19 mg, 0.188 mmol). After stirring for 16 h at room temperature,the reaction mixture is diluted with H₂O (10 mL) and extracted withEtOAc (50 mL). The organic phase is successively washed with 5% NaHCO₃aq, H₂O, and brine, then dried over Na₂SO₄. The organic phase isconcentrated in vacuo to give the crude residue, which is purified bySiO₂ column chromatography to give Intermediate 1.1 as a white amorphousmaterial; ES-MS: M+H=559; HPLC: _(c)t_(Ret)=4.23 min.

Intermediate 1.2

To a solution of Intermediate 1.3 (450 mg, 1.09 mmol) in THF/H₂O (5/5mL) is added LiOH.H₂O (84 mg, 2 mmol) at 0° C. After stirring for 1 h atthe same temperature, the reaction is quenched by 5% aqueous KHSO₄ (20mL) and extracted with EtOAc (200 mL). The organic phase is washed withH₂O and brine, then dried over Na₂SO₄. The organic phase is concentratedin vacuo to give Intermediate 1.2 as a white amorphous material; ES-MS:M+H=401: _(c)t_(Ret)=3.30.

Intermediate 1.3

Intermediate 1.3 is synthesized by coupling reaction of Intermediate 1.7with (3S,5R)-starting material-F analogously to the preparation ofIntermediate 1.1: ES-MS: M+H=415: _(c)t_(Ret)=3.69 min.

Intermediate 1.4

To a solution of Intermediate 1.5 (230 mg, 1.20 mmol) in CH₂Cl₂ (10 mL)are added Et₃SiH (728 mg, 6.26 mmol) and TFA (1.48 g, 12.9 mmol) at 0°C., then the mixture is stirred at room temperature. After stirring for15 h, the reaction mixture is concentrated in vacuo. The residue issuspended in 5% aqueous NaHCO₃ and extracted with CH₂Cl₂. The organicphase is washed with H₂O and brine, then dried over Na₂SO₄. The organicphase is concentrated in vacuo and the resulting residue is purified bySiO₂ column chromatography to give Intermediate 1.4 as a white amorphousmaterial; ES-MS: M+H=177; HPLC: _(c)t_(Ret)=2.24 min.

Intermediate 1.5

To a solution of Intermediate 1.6 (300 mg, 1.45 mmol) in THF (10 mL) isadded MeLi (1M in Et₂O, 7.25 mmol) at 0° C., then the mixture is stirredat room temperature. After stirring for 1 h, the reaction mixture iscooled down to 0° C. and quenched with 5% aqueous NaHCO₃ (50 mL). Thereaction mixture is then extracted with EtOAc (200 mL). The organicphase is washed with H₂O and brine, and then dried over Na₂SO₄. Theorganic phase is concentrated in vacuo to give the crude residue, whichis purified by SiO₂ column chromatography to give Intermediate 1.5 as awhite amorphous material; ES-MS: M+H=193; HPLC: _(c)t_(Ret)=1.74 min.

Intermediate 1.6

To a solution of 6-chloronicotinic acid ethyl ester (1 g, 5.4 mmol) inNMP (10 mL) are added cyclopropylamine (4.12 g, 72.2 mmol) and K₂CO₃(2.2 g, 16 mmol) at room temperature, then the mixture is stirred at 70°C. After stirring for 12 h, the reaction mixture is cooled down to roomtemperature and quenched with H₂O (100 mL). The reaction mixture isextracted with EtOAc (200 mL). The organic phase is washed with H₂O andbrine, then dried over Na₂SO₄. The organic phase is concentrated invacuo to give the crude residue, which is recrystallized fromn-hexane/Et₂O to give Intermediate 1.6 as a white amorphous material;ES-MS: M+H=207; HPLC: _(c)t_(Ret)=1.98 min.

Intermediate 1.7

Intermediate 1.7 is synthesized by deprotection of Intermediate 1.8analogously to the preparation of Example 1: ES-MS: M+H=146:_(B)t_(Ret)=1.32 min.

Intermediate 1.8

To a solution of Boc-D-Leucinol (277.9 mg, 1.278 mmol) in DMF (5 mL)under N₂ at 0° C. is added NaH (80.3 mg of 60 wt % in mineral oil, 2.00mmol). After stirring at the same temperature for a few min, EtI (0.122mL, 1.53 mmol) is added. The resulting solution is stirred at rt for 2h. The reaction is quenched with H₂O and the mixture is extracted withEtOAc, and dried over Na₂SO₄. Concentration under reduced pressure givesthe crude product. The crude product is purified by silica gelchromatography to afford Intermediate 1.8: ES-MS: M+H-Boc=146:_(B)t_(Ret)=2.11 min.

EXAMPLE 2

A solution of Intermediate 2.1 (194.2 mg, 0.33 mmol) in 4N HCl in EtOAc(3 mL) under N₂ is stirred at RT for 35 min. Concentration under thereduced pressure gives Example 2 as a white amorphous material: ES-MS:M+H=485: _(C)t_(Ret)=3.08 min.

Intermediate 2.1

To a solution of Intermediate 2.2 (153.8 mg, 0.356 mmol) in CH₂Cl₂ (5mL) under N₂ at RT are added EDCl.HCl (95 mg, 0.42 mmol) and HOAt (70mg, 0.51 mmol). The resulting solution is stirred at the sametemperature for 15 min. Then, a solution of Intermediate 2.4 (73 mg,0.35 mmol) and triethylamine (0.25 mL, 1.78 mmol) in CH₂Cl₂ (3 ml) isslowly added at 0° C. The solution is stirred at RT for 60 min.Concentration under reduced pressure gives the crude residue, which ispurified by silica gel chromatography to afford Intermediate 2.1 as awhite amorphous material; ES-MS: M=585; HPLC: _(C)t_(Ret)=4.78 min.

Intermediate 2.2

To a solution of Intermediate 2.3 (160 mg, 0.359 mmol) in THF/H₂O (5/5mL) is added LiOH.H₂O (29 mg, 0.691 mmol) at 0° C. After stirring for 1h at the same temperature, the reaction is quenched by 5% aqueous KHSO₄(20 mL) and extracted with CH₂Cl₂ (50 mL). The organic phase is washedwith H₂O and brine, then dried over Na₂SO₄. The organic phase isconcentrated in vacuo to give Intermediate 2.2 as a white amorphousmaterial; ES-MS: M+H=432; HPLC: _(c)t_(Ret)=3.50 min.

Intermediate 2.3

To a solution of (3R,5S)-starting material-F (293 mg, 1.02 mmol) in THF(10 mL) are added Et₃N (206 mg, 2.04 mmol) and TcBocCl (488 mg, 2.04mmol) at 0° C. After stirring for 1 h at the same temperature,MgBr₂.Et₂O (527 mg, 2.04 mmol) and Intermediate 1.4 (180 mg, 1.02 mmol)are added at 0° C., then the mixture is stirred at room temperature.After stirring for 3 h, the reaction is quenched with 5% aqueous KHSO₄(50 mL) and extracted with EtOAc (100 mL). The organic phase is washedwith H₂O and brine, and then dried over Na₂SO₄. The organic phase isconcentrated in vacuo and the resulting residue is purified by SiO₂column chromatography to give Intermediate 2.3 as a white amorphousmaterial; ES-MS: M+H=446; HPLC: _(c)t_(Ret)=3.86 min.

Intermediate 2.4

Intermediate 2.4 is synthesized by deprotection of Intermediate 2.5analogously to the preparation of Example 1. ES-MS: M+H=172:_(B)t_(Ret)=1.55 min.

Intermediate 2.5

To a solution of commercially available N-Boc-L-cyclohexylglycinol (499mg, 2.05 mmol) in DMF (8 mL) under N₂ at RT are added NaH (164 mg, 4.10mmol) and EtI (179 μL, 2.26 mmol) at 0° C. The reaction mixture isstirred at RT for 2 h. Then, H₂O is added to the resulting solution. Theaqueous phase is extracted with CH₂Cl₂. The combined organic phases aredried over Na₂SO₄. Concentration under reduced pressure, followed bypurification with silica gel column chromatography give Intermediate2.5: white amorphous material, ES-MS: M+H=272: _(B)t_(Ret)=2.46 min.

EXAMPLE 3

A mixture of Intermediate 3.1 (65 mg, 0.098 mmol) and 4M HCl in dioxane(2 mL) is stirred at room temperature. After stirring for 1 h, thereaction mixture is concentrated in vacuo to give Example 3: ES-MS:M+H=563: _(C)t_(Ret)=4.21 min.

Intermediate 3.1

To a solution of Intermediate 3.2 (60 mg, 0.112 mmol) and Intermediate1.7 (24 mg, 0.132 mmol) in CH₂Cl₂ (1 mL) are added EDCl.HCl (32 mg,0.168 mmol), HOAt (23 mg, 0.168 mmol), and Et₃N (17 mg, 0.168 mmol) atroom temperature; then the mixture is stirred at room temperature. Afterstirring for 16 h at the same temperature, the reaction mixture isdiluted with H₂O (10 mL) and extracted with EtOAc (50 mL). The organicphase is successively washed with 5% NaHCO₃ aq, H₂O and brine, and thendried over Na₂SO₄. The organic phase is concentrated in vacuo to givethe crude residue, which is purified by SiO₂ column chromatography togive Intermediate 3.1 as a white amorphous material; ES-MS: M+H=663;HPLC: _(c)t_(Ret)=4.83 min.

Intermediate 3.2

To a solution of Intermediate 3.3 (85 mg, 0.155 mmol) in THF/H₂O (5/5mL) is added LiOH.H₂O (14 mg, 0.330 mmol) at 0° C. After stirring for 1h at the same temperature, the reaction mixture is quenched with 5%aqueous KHSO₄ (20 mL) and extracted with Et₂O (100 mL). The organicphase is washed with H₂O and brine, and then dried over Na₂SO₄. Theorganic phase is concentrated in vacuo to give Intermediate 3.2 as awhite amorphous material; ES-MS: M+H=536; HPLC: _(c)t_(Ret)=4.32 min.

Intermediate 3.3

To a solution of (3R,5S)-starting material-F (103 mg, 0.36 mmol) inCH₃CN (1 mL) are added Et₃N (43 mg, 0.43 mmol), TcBocCl (86 mg, 0.36mmol) at 0° C. After stirring for 1 h at the same temperature,MgBr₂.Et₂O (112 mg, 0.43 mmol) and Intermediate 3.4 (100 mg, 0.36 mmol)are added at 00° C., then the mixture is stirred at room temperature.After stirring for 24 h, the reaction mixture is quenched with 5%aqueous KHSO₄ and extracted with EtOAc. The organic phase is washed withH₂O and brine, and then dried over Na₂SO₄. The organic phase isconcentrated in vacuo to give the crude residue, which is purified bySiO₂ column chromatography to give Intermediate 3.3 as a white amorphousmaterial; ES-MS: M+H=550; HPLC: _(c)t_(Ret)=4.65 min.

Intermediate 3.4

To a solution of Intermediate 3.5 (550 mg, 1.85 mmol) in CH₂Cl₂ (20 mL)are added Et₃SiH (2.3 g, 10.8 mmol) and TFA (7.4 g, 64.5 mmol) at 0° C.The resulting mixture is stirred at room temperature. After stirring for15 h, the reaction mixture is concentrated in vacuo. The residue issuspended in 5% aqueous NaHCO₃ and extracted with CH₂Cl₂. The organicphase is washed with H₂O and brine, and then dried over Na₂SO₄. Theorganic phase is concentrated in vacuo to give a crude residue, which ispurified by SiO₂ column chromatography to give Intermediate 3.4 as awhite amorphous material; ES-MS: M+H=281; HPLC: _(c)t_(Ret)=3.61 min.

Intermediate 3.5

To a solution of Intermediate 3.6 (574 mg, 1.7 mmol) in THF (18 mL) isadded MeMgBr (0.96 M solution in ether, 8.83 mL, 8.48 mmol) at 0° C. Theresulting mixture is stirred at room temperature for 1 h. After addingwater, the mixture is extracted with EtOAc, washed with NaHCO₃ aq.,brine and dried over MgSO₄. The organic phase is concentrated in vacuoto give the crude residue, which is purified by SiO₂ columnchromatography to give Intermediate 3.5 as a colorless oil; ES-MS:M+H=297; HPLC: _(c)t_(Ret)=3.50 min.

Intermediate 3.6

To a solution of Intermediate 3.7 (737 mg, 2.44 mmol) in NMP (17 mL) areadded K₂CO₃ (1.69 g, 12.2 mmol) and isobutylamine (535 mg, 7.32 mmol),and the resulting mixture is stirred at 120° C. overnight. After addingwater, the mixture is extracted with EtOAc, washed with water, brine anddried over MgSO₄. The organic phase is concentrated in vacuo to give thecrude residue, which is purified by SiO₂ column chromatography to giveIntermediate 3.6 as a colorless crystal; ES-MS: M+H=339; HPLC:_(c)t_(Ret)=3.94 min.

Intermediate 3.7

To a solution of Intermediate 3.8 (1 g, 4.07 mmol) in DMF (15 mL) areadded K₂CO₃ (1.69 g, 12.2 mmol) and isobutyliodide (1.5 g, 8.14 mmol),and the resulting mixture is stirred at 60° C. for 2 hr. The mixture isthen diluted with EtOAc, washed with water, sat. KHSO₄ aq., sat. NaHCO₃aq., brine and dried over MgSO₄. The organic phase is concentrated invacuo to give the crude residue, which is purified by SiO₂ columnchromatography to give Intermediate 3.7 as a colorless crystal; ES-MS:M+H=302; HPLC: _(c)t_(Ret)=4.49 min.

Intermediate 3.8

To a solution of 3-methoxypropan-1-ol (1.4 g) in THF (30 mL) is addedNaH (625 mg, 15.6 mmol) at 0° C. and the mixture is stirred for 30 minat r.t. Then, to the mixture is added a solution of4,6-dichloronicotinic acid (1.2 g, 6.25 mmol, U.S. Pat. 2005049419.) inTHF (10 mL) at 0° C. and it is next stirred for 3 h at r.t. After addingwater, the mixture is washed with ether. The aqueous phase is acidifiedwith KHSO₄ and then extracted with ether. The organic phase is washedwith brine and dried over MgSO₄. The organic phase is concentrated invacuo to give crude residue, which is purified by SiO₂ columnchromatography to give Intermediate 3.7 as a colorless crystal; ES-MS:M+H═; 246, HPLC: _(c)t_(Ret)=3.34 min.

EXAMPLE 4

Example 4 is synthesized by deprotection of Intermediate 4.1 (18 mg,0.03 mmol) analogously to the preparation of Example 1. Example 4:ES-MS: M+H=489: _(C)t_(Ret)=2.84 min.

Intermediate 4.1

A solution of Intermediate 4.2 (468 mg, 2.27 mmol), Intermediate 1.2 (1g, 2.5 mmol), BopCl (1.73 g, 6.81 mmol) and triethylamine (688 mg, 6.81mmol) is stirred at r.t. for 4 h. After adding sat. KHSO₄ aq., themixture is extracted with EtOAc. The organic layer is washed with water,sat. NaHCO₃ aq, brine and dried over MgSO₄. Silica gel columnchromatography gives Intermediate 4.1: colorless oil, ES-MS: M+H=589:_(B)t_(Ret)=1.92 min.

Intermediate 4.2

Intermediate 4.3 (2 g, 9 mmol) is dissolved in CH₂Cl₂ (14 ml). To thesolution is added triethylsilane (14 mL) and TFA (14 mL) at r.t. and themixture is stirred at 50° C. for 3 h. The solvent is removed undervacuum, and the residue is diluted with AcOEt. The resulting mixture iswashed with sat. NaHCO₃ aq., brine and dried over MgSO₄. Silica gelcolumn chromatography gives Intermediate 4.2: colorless crystal, ES-MS:M+H=207: _(B)t_(Ret)=1.51 min.

Intermediate 4.3

To a solution of Intermediate 4.4 (2.5 g, 2.63 mmol) in NMP (20 mL) isadded K₂CO₃ (9.2 g, 66.6 mmol) and isobutyl iodide (2.3 mL, 20 mmol).The resulting mixture is stirred at 80° C. for 40 min. Thencyclopropylamine (4.6 mL, 66.6 mmol) is added and the reaction mixtureis stirred overnight at 110° C. After adding water, the mixture isextracted with AcOEt. The organic layer is washed with water, brine anddried over MgSO₄. Recrystallization from EtOAc-n-hexane givesIntermediate 4.3: colorless crystal, ES-MS: M+H=265: _(B)t_(Ret)=1.54min.

Intermediate 4.4

To a suspension of NaH (5.2 g, 130 mmol) in THF (100 mL) is added MeOH(4.2 g, 130 mmol) at 0° C. After stirring at r.t. for 30 min, a solutionof 4,6-dichloronicotinic acid (10 g, 52.9 mmol, U.S. Pat. 2005049419.)in THF (100 mL) is added dropwise at 0° C. The resulting mixture isstirred at room temperature overnight. After adding water, the mixtureis washed with ether. The aqueous phase is acidified with KHSO₄ and thenextracted with ether. The organic phase is washed with brine and driedover MgSO₄. Concentration under reduced pressure gives Intermediate 4.4:colorless crystal, ES-MS: M+H=188: _(C)t_(Ret)=1.80 min.

EXAMPLE 5

Example 5 is synthesized by deprotection of Intermediate 5.1 analogouslyto the preparation of Example 1. Example 5: ES-MS: M+H=515:_(c)t_(Ret)=3.92 min.

Intermediate 5.1

Intermediate 5.1 is synthesized by coupling reaction of Intermediate 5.2with Intermediate 2.4 analogously to the preparation of Intermediate1.1: ES-MS: M+H=615: _(c)t_(Ret)=4.45 min.

Intermediate 5.2

Intermediate 5.2 is synthesized by saponification of Intermediate 5.3 (1g, 2.1 mmol) analogously to the preparation of Intermediate 1.2.Intermediate 5.2; colorless oil, ES-MS: M=462; HPLC: _(C)t_(Ret)=2.85min.

Intermediate 5.3

Intermediate 5.3 is synthesized by condensation of Intermediate 4.2 (800mg, 3.88 mmol) and (3R,5S)-Starting material-F (1230 mg, 4.27 mmol)analogously to the preparation of Intermediate 1.3. Intermediate 5.3;colorless oil, ES-MS: M=476; HPLC: _(C)t_(Ret)=1.78 min.

EXAMPLE 6

Example 6 is synthesized by deprotection of Intermediate 6.1 analogouslyto the preparation of Example 1. Example 6: ES-MS: M+H=505:_(c)t_(Ret)=3.29 min.

Intermediate 6.1

Intermediate 6.1 is synthesized by coupling reaction of Intermediate 6.2with Intermediate 1.7. analogously to the preparation of Intermediate1.1. Intermediate 6.1: ES-MS: M+H=605: _(c)t_(Ret)=4.11 min.

Intermediate 6.2

Intermediate 6.2 is synthesized by saponification of Intermediate 6.3analogously to the preparation of Intermediate 1.2. Intermediate 6.2:ES-MS: M+H=478: _(c)t_(Ret)=3.41 min.

Intermediate 6.3

Intermediate 6.3 is synthesized by coupling reaction of Intermediate 6.4with (3R,5S)-starting material-F analogously to the preparation ofIntermediate 1.3. Intermediate 6.3: S-MS: M+H=492: _(c)t_(Ret)=3.81 min.

Intermediate 6.4

Intermediate 6.4 is synthesized by dehydroxylation of Intermediate 6.5analogously to the preparation of Intermediate 1.4. Intermediate 6.4:ES-MS: M+H=223: _(c)t_(Ret)=2.78 min.

Intermediate 6.5

Intermediate 6.5 is synthesized by alkylation of Intermediate 6.6analogously to the preparation of Intermediate 1.5. Intermediate 6.5:ES-MS: M+H=239: _(c)t_(Ret)=1.95 min.

Intermediate 6.6

Intermediate 6.6 is synthesized from Intermediate 4.6 (usingisobutylamine instead of cyclopropylamine) analogously to thepreparation of Intermediate 4.5.

Intermediate 6.6: ES-MS: M+H=281: _(c)t_(Ret)=2.54 min.

EXAMPLE 7

Example 7 is synthesized by deprotection of Intermediate 7.1 analogouslyto the preparation of Example 1. Intermediate 7.1: ES-MS: M+H=445:_(c)t_(Ret)=3.44 min.

Intermediate 7.1

Intermediate 7.1 is synthesized by coupling reaction of Intermediate 2.2with Intermediate 7.2 analogously to the preparation of Intermediate1.1. Intermediate 7.1: ES-MS: M+H=545: _(c)t_(Ret)=4.30 min.

Intermediate 7.2

Intermediate 7.2 is synthesized by deprotection of Intermediate 7.3analogously to the preparation of Example 1. Intermediate 7.2: ES-MS:M+H=1³²:Bt_(Ret)=1.17 min.

Intermediate 7.3

Intermediate 7.3 is synthesized by alkylation of Boc-D-valinol [Journalof Organic Chemistry, 2000, 65 (16), 5037-5042] analogously to thepreparation of Example 1.8. Intermediate 7.3: ES-MS: M+H-Boc=132:_(B)t_(Ret)=2.03 min

EXAMPLE 8

Example 8 is synthesized by deprotection of Intermediate 8.1 analogouslyto the preparation of Example 1. Example 8: ES-MS: M+H=589:_(c)t_(Ret)=3.50 min.

Intermediate 8.1

To a solution of Intermediate 8.2 (49 mg, 0.239 mmol) and Intermediate1.2 (80 mg, 0.20 mmol) in CH₂Cl₂ are added BopCl (153 mg, 0.60 mmol) andEt₃N (61 mg, 0.60 mmol). After stirring for 25 h at room temperature,the reaction mixture is diluted with H₂O (10 mL) and extracted withEtOAc (50 mL). The organic phase is successively washed with 5% NaHCO₃aq, H₂O, and brine, then dried over Na₂SO₄. The organic phase isconcentrated in vacuo to give the crude residue, which is purified bySiO₂ column chromatography to give Intermediate 8.1 as a white amorphousmaterial; ES-MS: M+H=589; HPLC: _(C)t_(Ret)=3.50 min.

Intermediate 8.2

Intermediate 8.2 is synthesized by reduction of Intermediate 8.3analogously to the preparation of Intermediate 1.4. Intermediate 8.2:ES-MS: M+H=207: _(c)t_(Ret)=3.71 min.

Intermediate 8.3

To a solution of Intermediate 8.4 (500 mg, 2.5 mmol) in DMSO (10 mL) areadded cyclopropylamine (1.65 g, 28.9 mmol) and K₂CO₃ (415 mg, 7.5 mmol)at room temperature. The resulting mixture is then stirred at 80° C.After stirring for 5 h, the reaction mixture is cooled down to roomtemperature and quenched with H₂O (50 mL). The resulting mixture isextracted with EtOAc (100 mL). The organic phase is washed with H₂O andbrine, and then dried over Na₂SO₄. The organic phase is concentrated invacuo to give the crude residue, which is purified by SiO₂ columnchromatography to give Intermediate 8.3 as a yellow amorphous material;ES-MS: M+H=221; HPLC: _(c)t_(Ret)=2.34 min.

Intermediate 8.4

To a solution of Intermediate 8.5 (1.85 g, 7.56 mmol) in THF (40 mL) isadded MeLi (1M in Et₂O, 9.07 mmol) at 0° C., then the mixture is stirredat room temperature. After stirring for 1 h at room temperature, thereaction mixture is cooled down to 0° C. and quenched with 5% aqueousKHSO₄ (100 mL). The reaction mixture is then extracted with CH₂Cl₂ (100mL). The organic phase is washed with 5% aqueous NaHCO₃, H₂O and brine,and then dried over Na₂SO₄. The organic phase is concentrated in vacuoto give Intermediate 8.4 as a yellow amorphous material; ES-MS: M+H=200;HPLC: _(c)t_(Ret)=2.82 min.

Intermediate 8.5

To a solution of Intermediate 8.6 (2 g, 10 mmol) in ClCH₂CH₂Clare addedSOCl₂ (2 mL) and DMF (0.1 mL) at room temperature, then the mixture isstirred at 60° C. After stirring for 3 h at 60° C., the reaction mixtureis concentrated in vacuo to give a gummy material which is used in thenext reaction without further purification.

To a solution of the crude in CH₂Cl₂ is added N,O-dimethyl hydroxylaminehydrochloride (1.46 g, 15 mmol) and Et₃N (1.52 g, 15 mmol) at 0° C.,then the mixture is stirred at room temperature. After stirring for 19 hat room temperature, the reaction mixture is quenched with H₂O (100 mL)and extracted with CH₂Cl₂ (100 mL). The organic phase is successivelywashed with 5% NaHCO₃ aq, H₂O, and brine, and then dried over Na₂SO₄.The organic phase is concentrated in vacuo to give the crude residue,which is purified by SiO₂ column chromatography to give Intermediate 8.5as a brown oil; ES-MS: M+H=245; HPLC: _(c)t_(Ret)=2.27 min.

Intermediate 8.6

Intermediate 8.6 is synthesized by reaction of 2,4-dichloronicotinic(U.S. Pat. 2005049419.) acid with sodium ethoxide, analogously to thepreparation of Intermediate 4.6. Intermediate 8.6: ES-MS: M+H=202:_(c)t_(Ret)=2.12 min.

EXAMPLE 9

A solution of Intermediate 9.1 (13 mg, 0.02 mmol) in 4 NHCl-dioxane (4mL) under N₂ is stirred at 0° C. for 15 min. Then, the solution iswarmed up to room temperature and stirred at RT for 1 h. The mixture isconcentrated under reduced pressure to give Example 9 as a whiteamorphous material. ES-MS: M+H=563: _(A) t_(Ret)=2.48 min.

Intermediate 9.1

To a solution of Intermediate 9.2 (20 mg, 0.037 mmol) in CH₂Cl₂ (3 mL)under N₂ at 0° C. are added EDCl.HCl (11.6 mg, 0.075 mmol) and HOAt(10.1 mg, 0.075 mmol). After stirring at the same temperature for 5 min,Intermediate 1.7 (13.6 mg, 0.075 mmol) and Et₃N (0.04 mL, 0.22 mmol) areadded to the reaction mixture. The resulting solution is warmed up to RTand stirred at room temperature overnight. The reaction is quenched withH₂O and extracted with CH₂Cl₂. The combined organic extracts are washedwith H₂O and brine, dried over Na₂SO₄, filtered, and concentrated invacuo. The residue is purified by flash chromatography on silica gel togive the desired coupling product, Intermediate 9.1 as a white amorphousmaterial. ES-MS: M+H=663; HPLC: _(c)t_(Ret)=3.71 min.

Intermediate 9.2

To a solution of Intermediate 9.3 (20 mg, 0.04 mmol) in dry THF (1 mL)is added 0.22 M solution of LiOH (0.5 mL, 0.11 mmol) at 0° C. Theresulting solution is stirred at RT for 2 h. The reaction is quenchedwith saturated KHSO₄ solution (1 mL) and extracted with EtOAc. Upondrying over with Na₂SO₄, concentration under reduced pressure givesIntermediate 9.2 as a solid. This material is used in the next stepwithout further purification. ES-MS: M+H=536: _(c)t_(Ret)=2.93 min.

Intermediate 9.3

To a solution of (3R,4S,5S)-starting material-G (100 mg, 0.33 mmol) indry THF (5 mL) under N₂ is added(1-Chloro-2-methyl-propenyl)-dimethyl-amine (0.066 ml, 0.69 mmol) withcooling in an ice-bath. After stirring at the same temperature for 5min., the reaction is warmed up to RT and it is additionally stirred for20 min. The reaction mixture is concentrated under reduced pressure togive (3R,4R,5S)-5-Chlorocarbonyl-4-hydroxy-piperidine-1,3-dicarboxylicacid 1-tert-butyl ester 3-methyl ester. This material is used in thenext step without further purification. To a solution of Intermediate9.4 (87 mg, 0.33 mmol) and Et₃N (0.13 mL, 0.99 mmol) in dry THF (2 mL)is added dropwise a solution of the acyl chloride in THF (2 mL) at 0°C., and the resulting mixture is stirred for 2 h at RT. The mixture isquenched with sat. aq. NaHCO₃ and washed with water, dried over Na₂SO₄and concentrated under reduced pressure. The residue is purified byreversed phase chromatography to afford Intermediate 9.3 as an oil.ES-MS: M+H=550: _(c)t_(Ret)=3.74 min.

Intermediate 9.4

Intermediate 9.5 (4.3 g, 15.3 mmol) is dissolved in CH₂Cl₂ (51 mL). Tothe solution is added triethylsilane (24.4 mL, 153 mmol) and TFA (22.7mL, 306 mmol) at RT. and the mixture is stirred at 35° C. for 10 h. Thesolvent is removed under vacuum, and the residue is diluted with AcOEt.The organic phase is washed with sat. NaHCO₃ aq., brine and dried overNa₂SO₄. Silica gel column chromatography gives Intermediate 9.4:Colorless crystal, ES-MS: M+H=265: _(C)t_(Ret)=2.53 min.

Intermediate 9.5

Intermediate 9.6 (5.3 g, 16.4 mmol) is dissolved in THF (84 mL). To thesolution is added 0.97 M MeMgBr in THF (85 mL, 82.2 mmol) at 0° C. andthe mixture is stirred for 1 h at the same temperature. After addingwater, the mixture is extracted with AcOEt. The organic layer is washedwith brine and dried over Na₂SO₄. Silica gel column chromatography givesIntermediate 9.5: Colorless crystal, ES-MS: M+H=281: _(C)t_(Ret)=1.75min.

Intermediate 9.6

Intermediate 9.7 (22.1 g, 90.2 mmol) is dissolved in NMP (250 mL). Tothe solution is added K₂CO₃ (37.2 g, 270.6 mmol) and isobutyl iodide(15.5 mL, 135.3 mmol) and the mixture is stirred at 80° C. for 1 h, thencyclopropylamine (33.9 mL, 451.0 mmol) is added and the mixture isstirred for overnight at 110° C. After adding water, the mixture isextracted with AcOEt. The organic layer is washed with water, brine anddried over Na₂SO₄. Silica gel column chromatography gives Intermediate9.6: Colorless crystal, ES-MS: M+H=323: _(C)t_(Ret)=2.61 min.

Intermediate 9.7

4,6-Dichloronicotinic acid (20 g, 104.7 mmol, U.S. Pat. No.2,005,049419.) is dissolved in THF (120 mL). The solution is added to asolution of NaH (11.4, 262 mmol) and 3-methoxy-propan-1-ol (23.6 g, 262mmol) in THF (120 mL) at 0° C. and the resulting mixture is stirred for2 h at RT. After adding water, the mixture is washed with ether. Theaqueous layer is acidified with 1 N aq. HCl and extracted with EtOAc.The organic layer is washed with brine and dried over Na₂SO₄. Removal ofthe solvent under vacuum gives Intermediate 9.7: Colorless crystal,ES-MS: M+H=246: _(C)t_(Ret)=2.14 min.

EXAMPLE 10

A solution of Intermediate 10.1 (262 mg, 0.46 mmol) in 4N HCl in dioxane(3 mL) under N₂ is stirred at RT for 40 min. Concentration under thereduced pressure gives the HCl salt. Then, sat. NaHCO₃ aq. is added. Theaqueous phase is extracted with EtOAc. The combined organic extracts aredried over Na₂SO₄. Concentration under reduced pressure gives the cruderesidue, which is purified by silica gel chromatography followed bypreparative reversed phase HPLC to give the desired amine (170.2 mg,0.358 mmol). To a solution of amine in toluene (3 mL) is added 4NHCl-dioxane (3 mL). Concentration under reduced pressure gives thedesired Example 10 (HCl salt) as a white material: ES-MS: M+H=475:_(C)t_(Ret)=2.87 min.

Intermediate 10.1

To a solution of Intermediate 10.2 (174.5 mg, 0.39 mmol) in CH₂Cl₂ (10mL), under N₂ at RT, are added EDCl.HCl (130 mg, 0.57 mmol) and HOAt(100 mg, 0.73 mmol). The reaction mixture is stirred at the sametemperature for a few minutes. Then, Intermediate 1.7 (117 mg, 0.64mmol) and triethylamine (0.19 mL, 1.36 mmol) are added at 0° C. Theresulting solution is stirred at RT overnight, and then H₂O is added tothe reaction mixture. The aqueous phase is extracted with CH₂Cl₂. Thecombined organic phases are dried over Na₂SO₄. Concentration underreduced pressure gives the crude residue, which is purified by silicagel chromatography to afford Intermediate 10.1 as a white amorphousmaterial; ES-MS: M=575; HPLC: _(C)t_(Ret)=3.90 min.

Intermediate 10.2

To a solution of Intermediate 10.3 (180 mg, 0.39 mmol) in THF (7 mL) at0° C. is slowly added aqueous LiOH (40.7 mg, 0.97 mmol in H₂O (7 mL).The resulting solution is stirred at the same temperature for 25 min.The reaction mixture is diluted with H₂O and then washed with Et₂O. Theaqueous phase is acidified with sat. KHSO₄ aq. and then extracted withEt₂O. The combined organic extracts are dried over Na₂SO₄. Concentrationunder reduced pressure gives Intermediate 10.2 as a white amorphousmaterial; ES-MS: M=448; HPLC: _(C)t_(Ret)=3.00 min.

Intermediate 10.3

To a solution of (3R,4S,5S)-starting material-G (200 mg, 0.65 mmol) inCH₂Cl₂ (5 mL), under N₂ at 0° C., is added1-chloro-N,N,2-trimethyl-1-propenyl amine (0.1 mL, 0.76 mmol). Thesolution is stirred at the same temperature for 60 min. Then,Intermediate 1.4 (229 mg, 1.3 mmol) is added at 0° C. The resultingsolution is warmed to RT and stirred for 20 min, then H₂O is added andthe mixture is extracted with CH₂Cl₂. The combined organic extracts aredried over Na₂SO₄. Concentration under reduced pressure gives the cruderesidue, which is purified by silica gel chromatography to giveIntermediate 10.3. ES-MS: M=462; HPLC: _(C)t_(Ret)=3.66 min.

EXAMPLE 11

Example 11 is synthesized by deprotection of Intermediate 11.1analogously to the preparation of Example 1. Example 11: ES-MS: M+H=461:_(c)t_(Ret)=3.80 min.

Intermediate 11.1

Intermediate 11.1 is synthesized by coupling reaction of Intermediate10.2 with Intermediate 7.2 analogously to the preparation ofIntermediate 1.1. Intermediate 11.1: ES-MS: M+H=561: _(c)t_(Ret)=4.14min.

EXAMPLE 12

A solution of Intermediate 12.1 (9.3 mg, 0.015 mmol) in 4 NHCl-dioxane(1 mL), under N₂, is stirred at 0° C. for 15 min. Then, the solution iswarmed up to RT and stirred at room temperature for 20 min. The mixtureis concentrated under reduced pressure to give Example 12 as a whiteamorphous material. ES-MS: M+H=505: _(c)t_(Ret)=2.94 min.

Intermediate 12.1

To a solution of Intermediate 12.2 (63 mg, 0.13 mmol) in CH₂Cl₂ (3 mL)under N₂ at 0° C. are added EDCl.HCl (33.0 mg, 0.21 mmol) and HOAt (29.0mg, 0.21 mmol). After stirring at the same temperature for 5 min,Intermediate 1.7 (38.0 mg, 0.21 mmol) and Et₃N (0.06 mL, 0.42 mmol) areadded to the reaction mixture. The resulting solution is warmed up to RTand stirred at RT overnight. The reaction is quenched with H₂O andextracted with CH₂Cl₂. The combined organic extracts are washed with H₂Oand brine, dried over Na₂SO₄, filtered, and concentrated in vacuo. Theresidue is purified by flash chromatography on silica gel to giveIntermediate 12.1 as a white amorphous material. ES-MS: M+H=605; HPLC:_(c)t_(Ret)=3.93 min.

Intermediate 12.2

To a solution of Intermediate 12.3 (68 mg, 0.14 mmol) in dry THF (1 mL)is added 0.4 M solution of LiOH (1.0 mL, 0.41 mmol) at 0° C. Theresulting solution is stirred at RT for 2 h. The reaction is quenchedwith sat. KHSO₄ aq. (1 mL) and extracted with EtOAc. Upon drying overwith Na₂SO₄, concentration under reduced pressure gives Intermediate12.2 as a solid. This material is used in the next step without furtherpurification. ES-MS: M+H=478: _(A) t_(Ret)=2.77 min.

Intermediate 12.3

To a solution of (3R,4S,5S)-starting material-G (132 mg, 0.44 mmol) indry CH₂Cl₂ (2 mL), under N₂, is added(1-Chloro-2-methyl-propenyl)-dimethyl-amine (0.06 ml, 0.66 mmol) withcooling in an ice-bath. After stirring at the same temperature for 5min, the reaction is warmed up to RT and additionally stirred for 20min. The reaction mixture is concentrated under reduced pressure to give(3R,4R,5S)-5-Chlorocarbonyl-4-hydroxy-piperidine-1,3-dicarboxylic acid1-tert-butyl ester 3-methyl ester. This material is used in the nextstep without further purification. To a solution of Intermediate 12.4(90 mg, 0.44 mmol) and Et₃N (0.18 mL, 1.32 mmol) in dry CH₂Cl₂ (2 mL) isadded dropwise a solution of the acyl chloride in CH₂Cl₂ (2 mL) at 0° C.The resulting mixture is stirred for 2 h at room temperature. Themixture is quenched with sat. NaHCO₃ (3 mL) aq., washed with water,dried over Na₂SO₄, and concentrated under reduced pressure. The residueis purified by flash chromatography on silica gel to afford Intermediate12.3 as an oil. ES-MS: M+H=492: _(A)t_(Ret)=3.10 min.

Intermediate 12.4

In a 5 mL glass microwave vessel are placed Intermediate 1.5 (100 mg,0.52 mmol) and CAN (123.4 mg, 0.21 mmol) in dry MeOH (2 mL). Thereaction is heated under microwave irradiation using Biotage Initiator60 EXP at 10° C. for 15 min. The solvent is removed and the residue iswashed with sat. aq. NaHCO₃, water, dried over Na₂SO₄, and concentratedunder reduced pressure. The residue is purified by flash chromatographyon silica gel to give Intermediate 12.4 as a white solid material.ES-MS: M+H=207: _(c)t_(Ret)=3.02 min.

EXAMPLE 13

A solution of Intermediate 13.1 (107 mg, 0.177 mmol) in 4N HCl indioxane (3 mL) under N₂ is stirred at RT for 40 min. Concentration underreduced pressure gives the crude HCl salt. The crude is purified bypreparative reversed phase HPLC to give Example 13 as a white material:ES-MS: M+H=505: _(C)t_(Ret)=2.63 min.

Intermediate 13.1

To a solution of Intermediate 13.2 (122.7 mg, 0.257 mmol) in CH₂Cl₂ (5mL) under N₂ at RT are added EDCl.HCl (83 mg, 0.363 mmol) and HOAt (60.9mg, 0.447 mmol). The reaction mixture is stirred at the same temperaturefor a few minutes. Then, Intermediate 1.7 (69 mg, 0.38 mmol) andtriethylamine (0.11 mL, 0.82 mmol) are added at RT. The resultingsolution is stirred at the same temperature overnight. Concentrationunder reduced pressure gives crude residue, which is purified by silicagel chromatography to afford Intermediate 13.1 as a white amorphousmaterial; ES-MS: M=605; HPLC: _(C)t_(Ret)=3.39 min.

Intermediate 13.2

To a solution of Intermediate 13.3 (168 mg, 0.34 mmol) in THF (10 mL) at0° C. is slowly added aqueous LiOH (27.65 mg, 0.659 mmol in H₂O (10mL)). The resulting solution is stirred at the same temperature for 40min. The reaction mixture is diluted with H₂O and washed with Et₂O. Theaqueous phase is acidified with sat. KHSO₄ aq. and extracted with Et₂O.The combined organic extracts are dried over Na₂SO₄. Concentration underreduced pressure gives Intermediate 13.2 as a white amorphous material;ES-MS: M=478; HPLC: _(B)t_(Ret)=1.61 min.

Intermediate 13.3

To a solution of (3R,4S,5S)-starting material-G (499.7 mg, 1.647 mmol)in CH₂Cl₂ (33 mL) under N₂ at 0° C. is added1-chloro-N,N,2-trimethyl-1-propenyl amine (0.22 mL, 1.65 mmol). Thesolution is stirred at the same temperature for 60 min. Then,Intermediate 4.4 (683.3 mg, 3.3 mmol) is added at 0° C. The resultingsolution is warmed to RT and stirred for 60 min. Then, the mixture isdiluted with H₂O and extracted with CH₂Cl₂. The combined organicextracts are dried over Na₂SO₄. Concentration under reduced pressuregives the crude residue, which is purified by silica gel chromatographyto give Intermediate 13.3. ES-MS: M=492; HPLC: _(C)t_(Ret)=2.96 min.

EXAMPLE 14

A solution of Intermediate 14.1 (65 mg, 0.107 mmol) in 4N HCl in dioxane(2 mL) under N₂ is stirred at RT for 40 min. Concentration under reducedpressure gives the crude HCl salt. The crude is purified by preparativereversed phase HPLC to give Example 14 as a white material: ES-MS:M+H=505: _(C)t_(Ret)=2.79 min.

Intermediate 14.1

To a solution of Intermediate 14.2 (150 mg, 0.314 mmol) in CH₂Cl₂ (1 mL)under N₂ at RT are added EDCl.HCl (90 mg, 0.471 mmol) and HOAt (64 mg,0.471 mmol). The reaction mixture is stirred at the same temperature fora few minutes. Then, Intermediate 1.7 (69 mg, 0.38 mmol) andtriethylamine (0.066 mL, 0.471 mmol) are added at RT. The resultingsolution is stirred at the same temperature overnight. Concentrationunder reduced pressure gives crude residue, which is purified by silicagel chromatography to afford Intermediate 14.1 as a white amorphousmaterial; ES-MS: M=605; HPLC: _(C)t_(Ret)=3.56 min.

Intermediate 14.2

To a solution of Intermediate 14.3 (500 mg, 1.02 mmol) in THF (10 mL) at0° C. is slowly added aqueous LiOH (84 mg, 2.10 mmol in H₂O (10 mL). Theresulting solution is stirred at the same temperature for 40 min. Thereaction mixture is diluted with H₂O and washed with Et₂O. The aqueousphase is acidified with sat. KHSO₄ aq. and extracted with Et₂O. Thecombined organic extracts are dried over Na₂SO₄. Concentration underreduced pressure gives Intermediate 14.2 as a white amorphous material;ES-MS: M=478; HPLC: _(C)t_(Ret)=2.79 min.

Intermediate 14.3

To a solution of (3R,4S,5S)-starting material-G (1 g, 3.3 mmol) inCH₂Cl₂ (33 mL) under N₂ at 0° C. is added1-chloro-N,N,2-trimethyl-1-propenyl amine (0.523 mL, 3.95 mmol). Thesolution is stirred at the same temperature for 60 min. Then,Intermediate 8.2 (817 mg, 3.95 mmol) and Et₃N (0.552 mL, 3.95 mmol) areadded at 0° C. The resulting solution is warmed to RT and stirred for 60min. Then, the mixture is diluted with H₂O and extracted with CH₂Cl₂.The combined organic extracts are dried over Na₂SO₄. Concentration underreduced pressure gives the crude residue, which is purified by silicagel chromatography to give Intermediate 14.3. ES-MS: M=492; HPLC:_(C)t_(Ret)=3.01 min.

EXAMPLE 15

A solution of Intermediate 15.1 (15.1 mg, 0.023 mmol) in 4N HCl indioxane (3 mL) under N₂ is stirred at RT for 30 min. Concentration underreduced pressure gives Example 15 as a white material: ES-MS: M+H=565:_(C)t_(Ret)=4.30 min.

Intermediate 15.1

H₂ gas is streamed into a black suspension of intermediate 15.2 (22 mg,0.029 mmol) and 10% Pd—C (1 mg) in MeOH (2 mL) at RT. After stirring forovernight at RT, the reaction mixture is filtrated through Celite. Thefiltrate is concentrated under reduced pressure and purification withsilica gel column chromatography (hexane/EtOAc) give intermediate 15.1(15.1 mg, 0.023 mmol, 78%). ESI-MS (M+H): 665, HPLC: _(C)t_(Ret)=4.57min.

Intermediate 15.2

To a solution of intermediate 15.3 (34.9 mg) and intermediate 1.7 (12.1mg, 0.067 mmol) in CH₂Cl₂ (2 mL) are added EDCl.HCl (16 mg, 0.083 mmol),HOAt (11.3 mg, 0.083 mmol), and Et₃N (9.3 μL, 0.067 mmol) at roomtemperature, then the mixture is stirred for overnight at RT. Afteradding water, the mixture is extracted with CH₂Cl₂.

The organic layer is dried and concentration under reduced pressure andpurification with silica gel column chromatography (hexane/EtOAc) togive intermediate 15.2 (22 mg, 0.029 mmol).; ES-MS: M⁺=755:_(B)t_(Ret)=2.54 min.

Intermediate 15.3

To a solution of intermediate 15.4 (36.6 mg) in THF/H₂O (3/3 mL) isadded LiOH.H₂O (6.8 mg, 0.29 mmol) at 0° C. After stirring for 30 min atRT, the reaction is quenched with 5% KHSO₄ aq. And the mixture isextracted with Et₂O. The organic layer is dried over Na₂SO₄.Concentration under reduced pressure gives intermediate 15.3 (34.9 mg);ESI-MS (M+H): 628, HPLC: _(C)t_(Ret)=4.66 min.

Intermediate 15.4

To a solution of (3R,4S,5S)-starting material-G (100 mg, 0.33 mmol) inCH₂Cl₂ (2 mL) under N₂ at RT, 1-chloro-N,N-2-trimethyl-1-propenyl amine(55.7 μL, 0.42 mmol) is added. The solution is stirred at RT for 30 min.Then intermediate 15.5 (100 mg, 0.28 mmol) and trimethylamine (62.6 μL,0.45 mmol) in CH₂Cl₂ (2 mL) is added at 0° C. The resulting solution iswarmed to RT and is stirred at RT for 1 hr. The water and 1N HCl areadded. The aqueous phase is extracted with EtOAc. The organic layer iswashed with brine, and dried over Na₂SO₄. Concentration under reducedpressure and purification with preparative TLC (hexane/EtOAc) giveintermediate 15.4 (36.6 mg); ESI-MS (M+H): 642, HPLC: _(B)t_(Ret)=2.39min.

Intermediate 15.5

To a solution of intermediate 15.6 (801.1 mg, 2.15 mmol) in CH₂Cl₂ (3.5mL) is added triethylsilane (3.4 mL, 21.1 mmol) and TFA (3.3 mL, 43.0mmol) at 0° C. then the mixture is stirred at 40° C. for 3 hrs. Afterthe bulk of the solvent is concentrated in vacuo, the residue wasdiluted with EtOAc, neutralized with 1 N NaOH aq. and the mixture isextracted with EtOAc. The organic layer is dried over Na₂SO₄,concentrated in vacuo. The residue is purified by silica gel columnchromatography (hexane/EtOAc) to give intermediate 15.5 (651 mg, 1.83mmol, 85%). ¹H NMR (400 MHz, CDCl₃) δ 7.76 (s, 1H), 7.35-7.27 (m, 5H),5.81 (s, 1H), 4.53 (s, 2H), 4.41 (brs, 1H), 4.08 (dd, J=6.6, 6.0 Hz,2H), 3.67 (dd, J=6.1, 6.0 Hz, 2H), 3.04 (dd, J=6.6, 6.0 Hz, 2H),3.02-2.97 (m, 1H), 2.11 (quint, J=6.0 Hz, 2H), 1.87 (quint, J=6.6 Hz,1H), 1.18 (s, 3H), 1.16 (s, 3H), 0.99 (s, 3H), 0.97 (s, 3H); ESI-MS(M+H): 357, HPLC: _(C)t_(Ret)=3.60 min.

Intermediate 15.6

To a solution of intermediate 15.7 (1.28 g, 3.1 mmol) in THF (16 mL) isadded 1 M MeMgBr in THF (15.9 mL, 15.4 mmol) at 0° C. After stirred atthe same temperature for 1 hr, the mixture is warmed up to RT andstirred for overnight at RT. After adding water and sat NaHCO₃ aq., themixture is extracted with EtOAc. The organic layer is washed with brineand dried over Na₂SO₄, concentrated in vacuo. The residue is purified bysilica gel column chromatography (hexane/EtOAc) to give intermediate15.6 (1.06 g, 2.8 mmol, 92%). ¹H NMR (400 MHz, CDCl₃) δ7.91 (s, 1H),7.35-7.27 (m, 5H), 5.86 (s, 1H), 4.53 (s, 3H), 4.18 (dd, J=6.1, 6 Hz,2H), 3.72 (brs, 1H), 3.66 (dd, J=6.1, 6 Hz, 2H), 3.05 (dd, J=6.5, 6 Hz,2H), 2.14 (quint, J=6.0 Hz, 2H), 1.87 (quint, J=6.5 Hz, 1H), 1.56 (s,6H), 0.99 (s, 3H), 0.97 (s, 3H); ESI-MS (M+): 372, HPLC:_(C)t_(Ret)=3.12 min.

Intermediate 15.7

To a solution of intermediate 15.8 (1.56 g, 4.1 mmol) in NMP (42 mL) atRT is added K₂CO₃ (856 mg, 6.2 mmol) and isobutyl amine (604 μL, 6.2mmol) and the mixture is stirred for overnight at 110° C. After addingwater, the mixture is extracted with EtOAc. The organic layer is washedwith brine and dried over Na₂SO₄, concentrated in vacuo. The residue ispurified by silica gel column chromatography (hexane/EtOAc) to giveintermediate 15.7 (1.3 g, 3.1 mmol, 75%). ¹H NMR (400 MHz, CDCl₃) δ 8.60(s, 1H), 7.38-7.22 (m, 5H), 5.77 (s, 1H), 4.95-4.86 (m, 1H), 4.52 (s,2H), 4.15 (dd, J=6.1, 6.0 Hz, 2H), 4.00 (d, J=6.6 Hz, 2H), 3.71 (dd,J=6.1, 5.5 Hz, 2H), 3.09 (dd, J=6.6, 6.0 Hz, 2H), 2.19-2.10 (m, 2H),2.06-1.95 (m, 1H), 1.94-1.83 (m, 1H), 1.00 (s, 3H), 0.99 (s, 3H), 0.98(s, 3H), 0.97 (s, 3H); ESI-MS (M+): 414, HPLC: _(C)t_(Ret)=3.59 min.

Intermediate 15.8

The above crude Intermediate 15.9 (10.4 mmol) is dissolved in NMP (52mL). To the solution is added K₂CO₃ (4.32 g, 31.2 mmol) and isobutyliodide (1.8 mL, 15.6 mmol) and the mixture is stirred at 80° C. for 1hr. After adding water, the mixture is extracted with EtOAc. The organiclayer is washed with brine and dried over Na₂SO₄, concentrated in vacuo.The residue is purified by silica gel column chromatography(hexane/EtOAc) to give intermediate 15.8 (3.1 g, 8.2 mmol, 79%). ¹H NMR(400 MHz, CDCl₃) δ 8.71 (S, 1H), 7.36-7.23 (m, 5H), 6.91 (s, 1H), 4.51(s, 2H), 4.21 (t, J=6.0 Hz, 2H), 4.06 (d, J=7.0 Hz, 2H), 3.68 (t, J=6.0Hz, 2H), 2.20-2.10 (m, 2H), 2.08-1.97 (m, 1H), 1.00 (s, 3H), 0.98 (s,3H); ESI-MS (M+): 377, HPLC: _(B)t_(Ret)=2.31 min.

Intermediate 15.9

4,6-Dichloronicotinic acid (2 g, 10.4 mmol) is dissolved in THF (20 mL).The solution is added to a solution of NaH (1.04 g, 26.0 mmol) and3-benzyloxy-1-propanol (4.1 mL, 26.0 mmol) in THF (80 mL) at 0° C. andthe resulting mixture is stirred for 3 hrs at RT. After adding water,the mixture is acidified with 1 N HCl and then extracted with EtOAc. Theorganic extracts are dried over Na₂SO₄, concentrated in vacuo. Theresidue is purified by silica gel column chromatography (hexane/EtOAc)to give intermediate 15.9 as 3-benzyloxy-1-propanol mixture. ESI-MS(M+): 321, HPLC: _(B)t_(Ret)=1.82 min.

EXAMPLE 16

A solution of Intermediate 16.1 (65 mg, 0.116 mmol) in 4 N HCl indioxane (2 mL) under N₂ is stirred at RT for 2 h. Concentration underreduced pressure gives Example 16 as a white material: ES-MS: M+H=461:_(C)t_(Ret)=3.80 min.

Intermediate 16.1

To a solution of intermediate 10.2 (80 mg, 0.179 mmol) and intermediate7.2 (36 mg, 0.215 mmol) in CH₂Cl₂ (2 mL) are added EDCl.HCl (54 mg,0.281 mmol), HOAt (38 mg, 0.281 mmol), and Et₃N (28 mg, 0.281 mmol) atroom temperature, then the mixture is stirred at the same temperaturefor 2 h. The reaction is quenched by H₂O (10 mL) and extracted withEtOAc (50 mL). The organic extracts were washed with % NaHCO₃ aq, H₂O,and brine, then dried over Na₂SO₄. The organic phase was concentrated invacuo to give crude residue, which was purified by SiO₂ columnchromatography to give the Boc intermediate (65 mg, 0.116 mmol, 65%) asa colorless amorphous; ES-MS: M+H=561, HPLC: _(c)t_(Ret)=4.64 min.

EXAMPLE 17

Intermediate 17.1 (66 mg, 0.109 mmol) is dissolved in 4 NHCl-dioxane(0.5 mL) under N₂ at RT. After stirred for 15 min, concentration underthe reduced pressure gives the crude salt. Then, sat. NaHCO₃ aq. isadded. Organic phase is extracted with CH₂Cl₂, dried over Na₂SO₄.Filtration and concentration under the reduced pressure gives the crude.The crude is purified by silica gel chromatography to give Example 17(free base) (23.45 mg, 0.0466 mmol) in 43%. ES-MS: M+H=503,_(A)t_(ret)=2.10 min.

Intermediate 17.1

To a solution of intermediate 13.2 (91.6 mg, 0.192 mmol) in CH₂Cl₂ (3mL) under N₂, EDCl.HCl (60 mg, 0.26 mmol) and HOAt (49 mg, 0.36 mmol)are added at RT. After stirred at that temperature for a few minutes,intermediate 17.2 (34.3 mg, 0.19 mmol) and Et₃N (0.133 ml, 0.96 mmol),dissolved in CH₂Cl₂ (3 mL) are added. The resulting solution is stirredat that temperature for 1 hr. Concentration under the reduced pressuregives the crude. Purification by silica gel chromatography affordsintermediate 17.1 (66.0 mg, 0.109 mmol) in 57% yield. ES-MS: M+H=603,_(c)t_(ret)=3.21 min.

Intermediate 17.2

Intermediate 17.3 (46.8 mg, 0.192 mmol) is dissolved in 4 NHCl-dioxane(1 mL) under N₂. After stirred at RT for 30 min, concentration under thereduced pressure gives intermediate 17.2 (34.3 mg, 0.19 mmol) in 99%.ES-MS: M+H=144, _(c)t_(ret)=1.21 min. ¹H NMR (400 MHz, CDCl₃) δ 8.27(br, 2H), 7.90-7.55 (br, 1H), 3.31 (s, 3H), 3.18 (brd, J=6.04 Hz, 2H),3.13-3.00 (br, 1H), 2.40-2.00 (br, 3H), 1.88 (brd, J=13.12 Hz, 2H),1.70-1.40 (br, 2H), 1.18-0.99 (br, 2H).

Intermediate 17.3

To a solution of intermediate 17.4 (112.1 mg, 0.488 mmol) in THF (3 mL)under N₂, sodium hydride (19.5 mg in 60 wt % mineral oil, 0.488 mmol)and methyl iodine (30.3 uL, 0.488 mmol) are added at 0° C. The solutionis warmed to RT. After stirred at that temperature overnight, H₂O andsat. KHSO₄ aq. are added. Organic phase is extracted with ethyl acetate,dried over Na₂SO₄. Filtration and concentration under the reducedpressure gives the crude. Purification by silica gel chromatographyprovides the desired intermediate 17.3 (46.8 mg, 0.19 mmol) in 39%yield. ES-MS: M+H=244, _(c)t_(ret)=1.92 min. ¹H NMR (400 MHz, CDCl₃) δ4.40-4.30 (m, 1H), 3.37 (br, 1H), 3.32 (s, 3H), 3.17 (d, J=6.56 Hz, 2H),2.02 (d, J=9.56 Hz, 2H), 1.81 (d, J=11.6 Hz, 2H), 1.49 (m, 1H), 1.44 (s,9H), 1.13-0.98 (m, 4H).

Intermediate 17.4

To a solution of commercially availablecis-4-tert-butoxycarbonylamino-cyclohexanecarboxylic acid (200 mg, 0.82mmol) in THF (3 mL) under N₂, Et₃N (0.17 mL, 1.23 mmol) and isobutylchloroformate (0.106 mL, 0.82 mmol) are added at 0° C. After stirred atthat temperature for 30 min, filtration and concentration under thereduced pressure give anhydride.

To a solution of the crude in MeOH (5 mL) under N₂, NaBH₄ (170 mg, 4.49mmol) is added at 0° C. After stirred at that temperature for 40 min,sat. KHSO₄ aq. is added to the solution. Organic phase is extracted withCH₂Cl₂, dried over Na₂SO₄. Filtration and concentration under thereduced pressure give the crude. Purification by silica gelchromatography affords intermediate 17.4 (185.3 mg, 0.808 mmol) in 99%yield. ES-MS: M+H=230, _(c)t_(ret)=1.62 min. ¹H NMR (400 MHz, CDCl₃) δ4.38 (br, 1H), 3.45 (br, 2H), 3.38 (br, 1H), 2.04 (d, J=9.12 Hz, 2H),1.83 (d, J=10.8 Hz, 2H), 1.47 (s, 9H), 1.26 (dd, J=7.32, 7.08 Hz, 1H),1.15-0.99 (m, 4H).

EXAMPLE 18

A mixture of intermediate 18.1 form (180 mg, 0.305 mmol) and HCl indioxane (4 M solution, 2 mL) is stirred at room temperature for 1 h. Thereaction mixture is concentrated in vacuo to give example 18 (170 mg) asa colorless amorphous; ES-MS: M+H=491, HPLC: _(c)t_(Ret)=2.46 min.

Intermediate 18.1

To a solution of intermediate 14.2 (200 mg, 0.419 mmol) and intermediate7.2 (105 mg, 0.628 mmol) in CH₂Cl₂ (2 mL) are added EDCl.HCl (120 mg,0.628 mmol), HOAt (85 mg, 0.628 mmol), and Et₃N (64 mg, 0.628 mmol) atroom temperature, then the mixture is stirred at the same temperaturefor 2 h. The reaction is quenched by H₂O (50 mL) and extracted withEtOAc (100 mL). The organic extracts are washed with % NaHCO₃ aq, H₂O,and brine, then dried over Na₂SO₄. The organic phase is concentrated invacuo to give crude residue, which is purified by SiO₂ columnchromatography to give intermediate 18.1 (180 mg, 0.305 mmol, 73%) as acolorless amorphous; ES-MS: M+H=591, HPLC: _(c)t_(Ret)=3.37 min.

Biological Tests

Renin inhibitory activity was assessed in vitro by the method asoutlined above in item 2).

Results for Representative Compounds of Formula I

IC50 (FRET) Structure nM

0.9

3

0.6

0.4

0.9

2

0.9

0.3

2.5

0.8

2

0.6

0.8

0.3

6.0

2.5

9.0

0.6Equivalents

Those skilled in the art will recognize, or be able to ascertain usingno more than routine experimentation, numerous equivalents to thespecific procedures described herein. Such equivalents are considered tobe within the scope of the present invention and are covered by thefollowing claims. The appropriate components, processes, and methods ofthose patents, applications and other documents may be selected for thepresent invention and embodiments thereof.

We claim:
 1. A compound having a structure according to formula II

wherein R1 is C₁₋₇alkyl, which is optionally substituted by one, two orthree substituents selected from the group consisting of hydroxyl, haloand C₁-C₇-alkoxy; R2 is hydrogen, C₁₋₇alkyl, C₁-C₇-alkoxy, halo-C₁-C₇-alkyl, or halo-C₁-C₇-alkoxy; R3 is C₁₋₇alkyl or C₃₋₈cycloalkyl; and R5is C₁₋₇alkyl or C₃₋₈cycloalkyl; or a salt thereof.
 2. The compoundaccording to claim 1, wherein R1 is C₁₋₇alkyl, which is optionallysubstituted by C₁-C₇-alkoxy.
 3. The compound according to claim 1,wherein R2 is hydrogen or C₁-C₇-alkoxy.
 4. The compound according toclaim 1 wherein R3 is branched C₄₋₆ ₋alkyl or cyclopropyl.
 5. Thecompound according to claim 1, wherein R5 is C₁₋₄₋alkyl or cyclohexyl.6. The compound according to claim 1 selected from the compounds of theformula:

wherein R1, R2, R3 and R5 are R1 R2 R3 R5 1

H

2

H

4

5

6

7

H

8

or a pharmaceutically acceptable salt thereof, respectively.
 7. Apharmaceutical formulation, comprising: the compound according to claim1, or a pharmaceutically acceptable salt thereof, and at least onepharmaceutically acceptable carrier material.
 8. A method of treatinghypertension, wherein treating is alleviating or ameliorating at leastone symptom of said hypertension, comprising: administering to awarm-blooded animal in need of such treatment a pharmaceuticallyeffective amount of the compound according to claim 1 or apharmaceutically acceptable salt thereof.
 9. A process for themanufacture of the compound according to claim 1 said processcomprising: reacting a compound of the formula IV,

wherein PG is a protecting group; R4 is H and R5 is as defined in claim1, or an activated derivative thereof, with a compound of the formula V,

wherein R1, R2 and R3 are as defined in claim 1; or reacting a compoundof the formula VI,

wherein PG is a protecting group and R1, R2, R3 are as defined in claim1 and R4 is H, or an activated derivative thereof, with a compound ofthe formula VII,

wherein R5 is as defined in claim 1; and, if desired, subsequent to anyone or more of the processes mentioned above converting an obtainablefree compound of formula II into a salt thereof or converting a salt ofan obtainable compound of formula II into the free compound or adifferent salt; wherein any protecting groups are removed at anappropriate stage in order to obtain a corresponding compound of theformula II, or a salt thereof.
 10. A process for the manufacture of thecompound of Formula (VIII'a) or (VIII'b):

said process comprising reacting a compound of the formula VIII

wherein PG is a protecting group, with an alcohol R6OH, wherein R6 isunsubstituted or substituted alkyl or alkenyl, in the presence of achiral amine catalyst.
 11. A process for the manufacture of the compoundaccording to claim 1, said process comprising reacting a compound offormula (IXa)

wherein PG is a protecting group, R4 is H and R6 is unsubstituted orsubstituted alkyl or alkenyl, or an activated derivative thereof, with acompound of the formula V,

wherein R1, R2 and R3 are as defined in claim 1; to obtain the amide offormula Xa

wherein R4 is H; which is subjected to hydolysis of the ester moiety toobtain a compound of formula XIa

wherein R4 is H, which compound or an activated derivative thereof, isin turn reacted with a compound of the formula VII,

wherein R5 is as defined in claim 1, to obtain a compound of formula XII

and, if desired, subsequent to any one or more of the processesmentioned above converting an obtainable free compound of formula IIinto a salt thereof or converting a salt of an obtainable compound offormula II into the free compound or a different salt; wherein anyprotecting groups are removed at an appropriate stage in order to obtaina corresponding compound of the formula II, or a salt thereof.
 12. Aprocess for the manufacture of the compound according to claim 1, saidprocess comprising reacting a compound of formula (IXb)

wherein PG is a protecting group, R4 is H and R6 is unsubstituted orsubstituted alkyl or alkenyl, or an activated derivative thereof, with acompound of the formula VII,

wherein R5 is as defined in claim 1, to obtain the amide of formula Xb

which is subjected to hydrolysis of the ester moiety to obtain acompound of formula XIb

wherein R4 is H, which compound or an activated derivative thereof, isin turn reacted with a compound of the formula V,

wherein R1, R2 and R3 are as defined in claim 1, to obtain a compound offormula XII

and, if desired, subsequent to any one or more of the processesmentioned above converting an obtainable free compound of formula IIinto a salt thereof or converting a salt of an obtainable compound offormula II into the free compound or a different salt; wherein anyprotecting groups are removed at an appropriate stage in order to obtaina corresponding compound of the formula II, or a salt thereof.